Yazarlar TEM Connecticut Devlet kolej ve üniversite (CSCU) Merkezi nanoteknoloji ve Dr Ishita Mukerji Wesleyan Üniversitesi'nde için erişim için onu CD kullanarak yardım ve Dr. B. Ellen Scanley onun teknik eğitim için teşekkür etmek istiyorum Spektrofotometre. Raporlanan çalışma kısmen Bilim Enstitüsü Üniversitesi Fairfield, NASA Connecticut alan Grant Konsorsiyumu tarafından ve Ulusal Bilim Vakfı Hibe numarası CHE-1624774 altında tarafından desteklenmiştir.

1. sentez ve 1,2-Dithiolane arınma peptid modifiye
<ol><li>Sentezi dithiolane habercisi, 3-(acetylthio) -2-(acetylthiomethyl) propanoic asit19.
	<ol>
<li>3 1 g ekleyin-bromo - 2-(bromomethyl) propiyonik asit (1 eşdeğeri) 1 M NaOH (yaklaşık 4 mL) 55 ° C'de karıştırma ile 25 mL yuvarlak alt tepki şişesi için en az miktarda çözünmüş Reaksiyon şişeye bir septa ile mühür ve azot atmosferi altında yerleştirin.</li>
		<li>1,49 g potasyum thioacetate içeren bir çözüm hazırlamak (3.2 eşdeğeri) 4 ml deiyonize su ve 2 M sülfürik asit 3 mL (H2SO4) Tioasetik asit in situoluşturmak için.</li>
		<li>Tioasetik asit solüsyonu çek plastik tek kullanımlık 10 mL şırınga ve iğne şırıngayı yerleştirin. Karışımı dropwise septa iğneyle delme tarafından tepki şişeye ekleyin. Reaksiyon gecede 55 ° C'de devam</li>
		<li>Metanol ve diklorometan (1:9) karışımı kullanarak silika jel 60 F254 tabaklarda ince tabaka Kromatografi (TLC) tarafından tepki izlemek. Bromocresol yeşil leke tarafından reaksiyon ilerleme görselleştirin. Ürün bir Rf vardır 0,57 =.</li>
		<li>Tepki tam ve soğutmalı için oda sıcaklığında sonra pH 1 2 M H2karışıma çok acidify4. Sarı bir yağ dışarı çözüm ayırır.</li>
		<li>Soğuk kloroform (40 mL x 3) ile ürün ayıklayın. Organik katmanları birleştirmek ve magnezyum sülfat üzerinde kuru. Düşük basınç altında kloroform kaldırmak.</li>
		<li>İzole ürün, 1, kimliğini rakamlar 1B ve Cgösterildiği gibi nükleer manyetik rezonans (NMR) spektroskopisi ile onaylayın. Aşağıdaki sonuçlar bekliyoruz: 1H NMR, CDCl3, 300 MHz: d = 10.1 (b, 1 H), 3.2 (m, 4 H), (m, 1 H) 2,9, 2.4 (s, 6 H); 13 C NMR, CDCl3, 75 MHz: d = 195.1 (CH3COS-), 177.6 (-COOH), 45.1 (-CH2S-), 30.5 (CH), 29,2 (-SCOCH3). Not: Ürün sarı yağ ve % 83 genel bir verim vardır. Olmadan daha fazla arıtma ürün kullanın.</li>
	</ol></li>
	<li>SPP'ler ve Dithiolane habercisi-reçine kaplin Not: aşağıda açıklanan katı fazlı peptid sentez önerilen üreticinin iletişim kurallarını izleyen bir otomatik peptid synthesizer üzerinde yapılmıştır. Ayarları ve Kimyasalları uzman amino asitler kullanırken veya diğer ticari araçlar için adapte olabilir.<ol>
<li>Rink Amid 4-methylbenzhydrylamine (MBHA) reçine (0,1 mmol) 0,156 g tartmak ve bir reaksiyon gemi yerleştirin. Dimethylformamide (DMF) reçine en az 15 dk önce sentez başlangıcı için güzel.</li>
		<li>Her fluorenylmethyloxycarbonyl (Fmoc) 4 karşılıkları dışarı tartmak korumalı amino asit (0.4 mmol) dizisi ve 0,152 g N, N, N', N'- tetramethyl-O-(1H- benotriazol-1-yl) uranyum hexafluorophosphate (0.4 mmol, HBTU) için dizideki her amino asit. Her kartuş Fmoc korumalı amino asit ve HBTU içerir.</li>
		<li>Tüm ön sentez denetler synthesizer üzerinde çalıştırdıktan sonra (reaktifler, dolum reaktif satırları temizlenen ve tüm reaktif şişe basınçlı), c atlıkarınca amino asit kartuşları yerleştirin ' n ' terminus yön. Son amino asit pozisyon için son N-terminal Fmoc deprotection adım sonra bir boş kartuşu yerleştirin.</li>
		<li>Standart önerilen ayarlar kullanıldığında peptid sentez.<ol>
<li>Reçine ile 5 mL Fmoc grubundan, % 20 Piperodin DMF (5 dk x 2) içinde deprotect.</li>
			<li>DMF (6 x 5 mL) ile reçine kaplin adım önce yıkayın.</li>
			<li>Tek kaplin adım için 0.4 M N-methylmorpholine 4 mL DMF içinde Fmoc korumalı amino asit ve HBTU ekleyin. 30 amino asit Fmoc korumalı çözümü etkinleştirin çözüm tepki taşıyıcıyı aktarmadan önce s. Not: Otomatik peptid synthesizer reçine ve çözüm kabarcıklanma N2 gaz tarafından her 30 karışımları için 20 dk kaplin reaksiyon sırasında yer alır. El ile peptid sentez için tepki gemi bir orbital çalkalayıcı düşük hızda bağlantı adım süresince yerleştirin.</li>
			<li>Çözüm drenaj ve reçine DMF (3 x 5 mL) ile yıkayın.</li>
			<li>Her Fmoc korumalı amino asit c için adımları 1.2.4.1 1.2.4.4 aracılığıyla yineleyin ' n '-peptid ilgi sentezlemek için terminus yön.</li>
		</ol>
</li>
		<li>Son N-terminal deprotection adım sonra reçine tek kullanımlık fritted enjektör aktarın. Reçine DMF (3 x 5 mL) ve diklorometan (DCM, 3 x 5 mL) ile yıkayın. Not: Bir vakum desiccator yıkama DCM sonra reçine depolanabilir. Reçine kaplin önceki depolanmışsa, DMF reçine kaplin reaksiyon önce şişmeye emin olun.</li>
		<li>Dithiolane öncül (1)-reçine peptid N-terminus 1, 5 mL DMF, de HBTU 4 karşılıkları ve n, N-diisopropylethylamine (DIPEA) 10 eşdeğerleri 4 karşılıkları ekleyerek birkaç. 10 dk kaplin karışımı fritted şırınga içeren reçine eklemeden önce önceden etkinleştirin.</li>
		<li>Kaplin reaksiyon 2s için salla. 2 h sonra reçine DMF (3 x 5 mL) ile yıkayın ve bir gecede sallayarak ile kaplin tepki tekrarlayın.</li>
		<li>Sonra ikinci bağlantı, reçine DMF (3 x 5 mL) ve DCM (3 x 5 mL) ile yıkayın. Not: Reçine vakum altında bu noktada bölünme kadar saklanabilir.</li>
	</ol></li>
	<li>Thioacetate Deprotection ve peptid bölünme reçine üzerinden
	<ol>
<li>N-terminus dithiolane habercisi thioacetate grubundan deprotect için kuru reçine bir 10 mL mikrodalga tepki tüp aktarmak ve DMF 2 mL ekleyin. Reçine şişmeye, gemi küçük manyetik heyecan çubuğu eklemek ve bir alçak hız-in manyetik karıştırarak 15 dakika ile yeniden askıya sağlar.</li>
		<li>2 mL konsantre amonyum hidroksit ekleyin, reaksiyon teknesi ile silikon septa kap ve reaksiyon taşıyıcıyı karıştırma ile 45 dk için 75 ° C mikrodalga ayarlarını kullanarak bir mikrodalga reaktör içine koyun.</li>
		<li>Mikrodalga Reaksiyon tamamlandıktan sonra reçine temiz tek kullanımlık fritted enjektör aktarın. DMF (2 x 5 mL) ve metanol (MeOH, 2 x 5 mL) ile yıkayın.</li>
		<li>Metanol (1:4), konsantre amonyum hidroksit çözeltisi toplam hacmi 5 mL için ekleyin. Dithiolane ringde disülfür bağ intramolecular oksidasyon artırmak için gecede sallamak için bırakın.</li>
		<li>Reçine MeOH (2 x 5 mL) ve DCM (3 x 5 mL) ile yıkayın. Not: Kurutulmuş reçine bir vakum desiccator bu noktada depolanabilir.</li>
		<li>Bölünme kokteyl ile 1,5 saat için hafifçe sallayarak şırınga içeren reçine ekleyin. Göğüs arası kokteyl kullanılan % 95 trifluoroacetic asit (TFA), % 2.5 triisopropylsilane (ipuçları) ve toplam hacmi 5 mL % 2.5 su var. Dikkat: sadece kimyasal duman başlık altında çalışır. TFA uçucu ve aşındırıcı. Not: Yukarıdaki bölünme kokteyl çözüm peptid dizileri ve amino asit sidechain grupları korumak çoğunluğu için yeterlidir; Ancak, alternatif bölünme kokteyller için belirli amino asit yan zinciri gruplarında (belirli peptidler Cys, Met, Trp ve Arg içeren) veya diğer reçine Kimya20korunması gerekli olabilir.</li>
		<li>Dropwise ek fritted şırıngadan tarafından 25 mL 50 mL konik tüp içinde soğuk dietil eter içine ham peptid çökelti. Peptid beyaz katı precipitates. Pelet peptid Santrifüjü 1300 x g 10 dakika süreyle de tarafından özenle dikkatle boşaltmak dietil eter atık toplama için ayrı bir kaba.</li>
		<li>Başka bir 25 mL dietil eter konik tüp ekleyin ve çökelti vortexing tarafından yeniden askıya alma. Santrifüjü 1300 x g de 10 dakikadır tekrar ve tekrar dietil eter dikkatle boşaltmak. Pelet vakum altında kuru.</li>
	</ol></li>
	<li>1,2-Dithiolane arınma peptid modifiye Not: ham peptid ters faz-HPLC tarafından arındırmak. Toplamak ve peptid doruklarına birleştirmek ve kitle tarafından MALDI-TOF Kütle spektrometresi onaylayın.<ol>
<li>Asetonitril % 0.1 ile en az miktarda ham peptid Pelet dağıtılması TFA. Peptid'ın hydrophobicity ve toplama eğilimi nedeniyle örnek 40 ° C'de çözünürlük içinde yardım etmek için ısı yavaşça. Not: potansiyel disülfür Satım reaksiyonlar21,22,23önlemek için daha yüksek sıcaklık ve peptid sonication kaçının.</li>
		<li>Ham peptid 1 mL HPLC arıtma için hazırlamak için konsantre peptid stokunun 400 μL Asetonitril içinde H2O %0.1 TFA ve filtre ile 600 μL 22 mikron şırınga filtreden HPLC şişe ekleyin. Ek bir % 5 isopropanol peptid toplama ve yağış önlemek için ilave edilebilir.</li>
		<li>3 mL/dk 15-%55 Asetonitril 20 dak Set doğrusal bir gradyan üzerinde bir akış hızı ile C-18 yarı partiye hazırlık sütun kullanarak peptid 222 için UV dedektörleri arındırmak nm (Amid omurga) ve 330 nm (disülfür bağ). Toplamak ve faiz (Şekil 2A) doruklarına birleştirmek.</li>
		<li>Peptid ürün kitle tarafından MALDI-TOF Kütle Spektrometre reflektron modunda (Şekil 2B) onaylayın. Analiz için 0.5 μL maldı plaka üzerinde toplanan Peak 2,5-dihidroksibenzoik asit (DHB) matris 0.5 μL ile karıştırın (10 mg/mL DHB % 50 Asetonitril, % 0,1 TFA). Not: Ortak adducts MALDI-TOF Kütle spektrometresi dahil sodyum ve potasyum tuzu adduct ([M + Na]+ ve [M + K]+) tepeler. Örnek analiz önce desalting tavsiye edilir Eğer tuz adduct doruklarına ana [M + H]+ en yüksek sinyal bastırmak. Ayrıca, [M + O]+ oksitlenmiş bir zirve da tespit 1,2-dithiolane değiştiren peptid. Lazer indüklenen oksidasyon DHB matrisi kullanarak maldı iyonlaşma gelen raporda örnek toplama, solvent ve lazer şiddeti gibi faktörlere bağlı maldı oksidasyon artifakı24sınırlamak için değiştirilebilir öneriyor.</li>
		<li>Onay alındıktan sonra MALDI-TOF'doğru kütlesinin peptid sonra flaş dondurma lyophilize. Peptid liyofilize toz vakum altında derleme kadar tutun.</li>
	</ol></li>
</ol>2. karakterizasyonu Supramolecular kendinden montajlı yapıları
<ol><li>
Amiloid lifleri oluşumu
	<ol>
<li>Kendinden montajlı çözüm hazırlamak için bir analitik denge kullanarak peptid toz 1 mg tartın. (PH 7.5) % 20 Asetonitril ve karışımı 10 mM (4-(2-hydroxyethyl)-1-piperazineethanesulfonic asit (HEPES) son konsantrasyonu 1 mg/mL peptid derleme karışımı bir 1.5 mL microcentrifuge tüp. içine dağıtılması Girdap derleme çözüm ve oda sıcaklığında bir araya bırakın.</li>
	</ol>
</li>
	<li>
Amiloid liflerinin spektroskopik karakterizasyonu
	<ol>
<li>Peptid derleme işlemi birkaç günde Fourier dönüşümü kızılötesi (FTIR) spektroskopisi ile izleyin. Örnek17unassembled peptidler kaynaklanan IR imza, 1670 cm-1 merkezli bir geniş tepedir. Peptid montaj örnekleri genellikle kaybolur ve olgunlaşma ulaşmak geniş unassembled zirve için bir iki hafta sürebilir.<ol>
<li>ATR elmas kristal üzerinde ince bir film olarak 8-10 μL derleme çözümün bir aliquot kuru. 1640 1630 cm-1 bir büyük ve geniş su tepe kaybolması Kuru film formları olarak izlemek.</li>
			<li>IR spectra 2 cm-1 çözünürlük 50 taramaları ortalama 1500-1800 cm-1 üzerinden elde etmek. Elde etmek ve arka plan inceden inceye gözden geçirmek her örnek taramadan önce çıkarma. Cm-1 (Şekil 3A)25,261625 ve 1635 arasında keskin bir tepe β-yapraklık derleme için IR imzadır.</li>
		</ol>
</li>
		<li>β-yapraklık zengin supramolecular yapıları peptid derlemesini dairesel dichroism (CD) tarafından karakterize. CD spectropolarimeter ile Peltier bir sıcaklık kontrol sistemi kullanarak spectra kaydedin.<ol>
<li>0,1 mm yol uzunluğu microcuvette içinde derleme çözümün 30 μL pipet. Not: Bir hücre tutucu klamp ve araç kısa yol uzunluğu hücreye yerleştirin için gereklidir.</li>
			<li>Her spektrum için CD enstrüman için aşağıdaki parametreleri ayarlayın: 300 dalga boylarında tarama nm 180 nm, tarama hızı 100/dk, bant genişliği 1 nm, 25 ° C, ortalama üç inceden inceye gözden geçirmek için.</li>
			<li>Arabellek (% 20 Asetonitril/10 mM HEPES, pH 7.5) bir spektrum toplamak ve her örnek tarama denetimi olarak değerden çıkarmak. β-yaprak CD imzası şöyledir: bir ellipticity en az yaklaşık 220 nm (Şekil 3B)27merkezli.</li>
		</ol>
</li>
	</ol>
</li>
	<li>
Amiloid liflerinin mikroskobu
	<ol>
<li>2-3 hafta β-yapraklık zengin supramolecular yapılarına olgunlaşmaya peptid örnekleri için izin. Not: Derlemelerin derleme işlemi de daha önceki aşamalarında transmisyon elektron mikroskobu (TEM) kullanarak yansıma.<ol>
<li>Peptid derleme çözüm TEM karbon kılavuz yüzeyinin 10 μL pipet. Not: pipet ucu ızgara yüzeyine dokunmamaya dikkat çekmek. Yüksek hassasiyetli, kendi kendine kapanış cımbız TEM kılavuz hazırlık sırasında tutmak için kullanılır.</li>
			<li>Derlemeler ızgara yüzeyine absorbe izin vermek için 1-2 dakika bekleyin. Aşırı örnek filtre kağıdı kılavuz kenarına dokunarak kaldırın.</li>
			<li>% 2 uranyl asetat leke piyasada bulunan %4 uranyl asetat eriyik-e doğru 100 μL deiyonize su ekleyerek hazırlayın. % 2 uranyl asetat leke kılavuz yüzeyine 10 μL pipet ve 2-3 dakika için kuluçkaya. Kuluçka sonra fazla leke kılavuz kenarına filtre kağıdı dokunarak kaldırın.</li>
			<li>TEM Izgaralar bir vakum desiccator gecede yerleştirin. Vakum altında görüntüleme kadar saklayın.</li>
			<li>TEM (Şekil 3 c) ile hazırlanmış örnekler görüntü. Mikroskopi için tipik parametreleri aşağıdaki gibidir: 9.300 X 23 için değişen büyüklüklerde görüntülere 000 X, tungsten filaman 120 hızlanan bir gerilim ile kV. Not: ImageJ edinsel TEM görüntüleri28supramolecular yapılarının ortalama lif genişliğini ölçmek için kullanılabilir. Dikkat: Tüm ilgili güvenlik bilgi formları (SDS) kullanmadan önce lütfen danışın. Çeşitli kimyasalların sentezinde, arıtma, kullanılan ve açıklanan 1,2-dithiolane değiştiren Self montaj peptidler karakterizasyonu aşındırıcı veya toksik ve yalnızca bir kimyasal duman başlık altında kullanılmalıdır. Her zaman uygun kişisel koruyucu ekipman (koruyucu gözlük, laboratuvar önlüğü, tam uzunlukta pantolon, kapalı-toe Ayakkabı dahil olmak üzere) kullan laboratuvar olarak çalışırken.</li>
		</ol>
</li>
	</ol>
</li>
</ol>

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L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Polymerized+liposomes+formed+under+extremely+mild+conditions.">Polymerized liposomes formed under extremely mild conditions.</a> Journal of the American Chemical Society. 108, 7789-7791 (1986).</li><li>Zhang, N., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=ATN-161+Peptide+functionalized+reversibly+cross-linked+polymersomes+mediate+targeted+doxorubicin+delivery+into+melanoma-bearing+C57BL/6+mice.">ATN-161 Peptide functionalized reversibly cross-linked polymersomes mediate targeted doxorubicin delivery into melanoma-bearing C57BL/6 mice.</a> Molecular Pharmaceutics. 14, 2538-2547 (2017).</li><li>Margulis, K., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Formation+of+polymeric+nanocubes+by+self-assembly+and+crystallization+of+dithiolane-containing+triblock+copolymers.">Formation of polymeric nanocubes by self-assembly and crystallization of dithiolane-containing triblock copolymers.</a> Angewandte Chemie International Edition. 56, 16357-16362 (2017).</li><li>Zhang, X., Waymouth, R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=1,2-Dithiolane-Derived+Dynamic,+Covalent+Materials:+Cooperative+Self-Assembly+and+Reversible+Cross-Linking.">1,2-Dithiolane-Derived Dynamic, Covalent Materials: Cooperative Self-Assembly and Reversible Cross-Linking.</a> Journal of the American Chemical Society. 139, 3822-3833 (2017).</li><li>Sakia, N., Matile, S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Stack+exchange+strategies+for+the+synthesis+of+covalent+double-channel+photosystems+by+self-organizing+surface-initiated+polymerization.">Stack exchange strategies for the synthesis of covalent double-channel photosystems by self-organizing surface-initiated polymerization.</a> Journal of the American Chemical Society. 133, 18542-18545 (2011).</li><li>Uji, H., Morita, T., Kimura, S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Molecular+direction+dependence+of+single-molecule+conductance+of+a+helical+peptide+in+molecular+junction.">Molecular direction dependence of single-molecule conductance of a helical peptide in molecular junction.</a> Physical Chemistry Chemical Physics. 15, 757-760 (2013).</li><li>Liang, C., Ni, R., Smith, J. E., Childers, W. S., Mehta, A. K., Lynn, D. G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Kinetic+intermediates+in+amyloid+assembly.">Kinetic intermediates in amyloid assembly.</a> Journal of the American Chemical Society. 136, 15116-15149 (2014).</li><li>Smith, J. E., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Defining+the+dynamic+conformational+network+of+cross-&#946;+peptide+assembly.">Defining the dynamic conformational network of cross-&#946; peptide assembly.</a> Israel Journal of Chemistry. 55, 763-769 (2015).</li><li>Black, S. P., Sanders, J. K. M., Stefankiewicz, A. R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Disulfide+exchange:+Exposing+supramolecular+reactivity+through+dynamic+covalent+chemistry.">Disulfide exchange: Exposing supramolecular reactivity through dynamic covalent chemistry.</a> Chemical Society Reviews. 43, 1861-1872 (2014).</li><li>Vendetti, A., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Dihydroasparagusic+acid:+Antioxidant+and+tyrosinase+inhibitory+activities+and+improved+synthesis.">Dihydroasparagusic acid: Antioxidant and tyrosinase inhibitory activities and improved synthesis.</a> Journal of Agricultural and Food Chemistry. 61, 6848-6855 (2013).</li><li>Stawikowski, M., Fields, G. B. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Introduction+to+peptide+synthesis.">Introduction to peptide synthesis.</a> Current Protocols in Protein Science. 26, (2002).</li><li>Canadell, J., Goossens, H., Klumperman, B. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Self-healing+materials+based+on+disulfide+links.">Self-healing materials based on disulfide links.</a> Macromolecules. 44, 2536-2541 (2011).</li><li>Lafont, U., van Zeijl, H., van der Zwaag, S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Influence+of+cross-linkers+on+the+cohesive+and+adhesive+self-healing+ability+of+polydisulfide-based+thermosets.">Influence of cross-linkers on the cohesive and adhesive self-healing ability of polydisulfide-based thermosets.</a> ACS Applied Materials and Interfaces. 4, 6280-6288 (2012).</li><li>Komaromy, D., Stuart, M. C. A., Santiago, G. M., Tezcan, M., Krasnikov, V. V., Otto, S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Self-assembly+can+direct+dynamic+covalent+bond+formation+toward+diversity+or+specificity.">Self-assembly can direct dynamic covalent bond formation toward diversity or specificity.</a> Journal of the American Chemical Society. 139, 6234-6241 (2017).</li><li>McAvery, K. M., Guan, B., Fortier, C. A., Tarr, M. A., Cole, R. B. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Laser-induced+oxidation+of+cholesterol+observed+during+MALDI-TOF+mass+spectrometry.">Laser-induced oxidation of cholesterol observed during MALDI-TOF mass spectrometry.</a> Journal of the American Society for Mass Spectrometry. 22, 659-669 (2011).</li><li>Krimm, S., Bandekar, J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Vibrational+spectroscopy+and+conformation+of+peptides,+polypeptides,+and+proteins.">Vibrational spectroscopy and conformation of peptides, polypeptides, and proteins.</a> Advances in Protein Chemistry. 38, 181-364 (1986).</li><li>Halverson, K. J., Sucholeiki, I., Ashburn, T. T., Lansbury, P. T. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Location+of+&#946;-sheet-forming+sequences+in+amyloid+proteins+by+FTIR.">Location of &#946;-sheet-forming sequences in amyloid proteins by FTIR.</a> Journal of the American Chemical Society. 113, 6701-6703 (1991).</li><li>Greenfield, N., Fasman, G. D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Computed+circular+dichroism+spectra+for+the+evaluation+of+protein+confirmation.">Computed circular dichroism spectra for the evaluation of protein confirmation.</a> Biochemistry. 8, 4108-4116 (1969).</li><li>. ImageJ Available from: <a target="_blank" href="https://imagej.nih.gov/ij">https://imagej.nih.gov/ij</a> (2016)</li><li>Roy, S., Shinde, S., Hamilton, G. A., Hartnett, H. E., Jones, A. K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Artificial+[FeFe]-hydrogenase:+On+resin+modification+of+an+amino+acid+to+anchor+a+hexacarbonyldiiron+cluster+in+a+peptide+framework.">Artificial [FeFe]-hydrogenase: On resin modification of an amino acid to anchor a hexacarbonyldiiron cluster in a peptide framework.</a> European Journal of Inorganic Chemistry. 2011, 1050-1055 (2011).</li><li>Van Duinen, S. G., Castano, E. M., Prelli, F., Bots, G. T. A. B., Luyendijk, W., Frangione, B. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Hereditary+cerebral+hemorrhage+with+amyloidosis+in+patients+of+Dutch+origin+is+related+to+Alzheimer+disease.">Hereditary cerebral hemorrhage with amyloidosis in patients of Dutch origin is related to Alzheimer disease.</a> Proceedings of the National Academy of Sciences of the United States of America. 84, 5991-5994 (1987).</li><li>Barth, A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+infrared+absorption+of+amino+acid+sidechains.">The infrared absorption of amino acid sidechains.</a> Progress in Biophysics and Molecular Biology. 74, 141-173 (2000).</li><li>Jayaraman, M., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Slow+amyloid+nucleation+via+&#945;-helix-rich+oligomeric+intermediates+in+short+polyglutamine-containing+Huntingtin+fragments.">Slow amyloid nucleation via &#945;-helix-rich oligomeric intermediates in short polyglutamine-containing Huntingtin fragments.</a> Journal of Molecular Biology. 415, 881-899 (2012).</li></ol>

Yazarlar ifşa gerek yok.

Bu makalede bir N-terminal 1,2-dithiolane değiştiren Self montaj peptid saflaştırılması ve karakterizasyonu elde edilen supramolecular yapıları ve hem sentezi ayrıntılarını anlatılır. Rapor burada 1,2-dithiolane peptid sentez 3-(acetylthio) -2 dithiolane habercisi üretmek için bir tek adımlı sentez de dahil olmak üzere avantajı vardır-(acetylthiomethyl) propanoic asit ve üzerinde reçine mikrodalga deprotection tepki Grup olarak toksik hidrazin deprotection için daha güvenli bir alternatif daha ö...

Kimya şekillendirme sağlam peptit bağı katı fazlı peptid sentezinde yer ve sıra uzunluğu ve kompozisyon kontrol yeteneği yapmak supramolecular yapılarına ağır araştırılmış bir alanı kendi kendine araya peptidler. Kontrol ve yan zincir steric dahil olmak üzere peptid kendi kendine monte yapılar ve elektrostatik etkileşimler, hidrojen bağı ve hidrofobik etkileri1, stabilize faktörler tasarım kuralları kümesi olarak hizmet vermektedir. Bu temel tasarım kuralları içine araştırma ilerlemeye devam ediyor gibi peptid mantıksal sonraki adımda kendinden montajlı çeşitliliği peptid temel yapıları ve işlevleri genişletme kapsar. Kendi kendine peptidler montaj sırasında birçok biyomedikal uygulamalarda peptit dizisi veya derleme koşullar2,3,4, için strateji geliştirme ayarlama tarafından kullanılan çok yönlü bir biomaterial vardır peptid nanofibers5,6,7,8,9 sonrası derleme değişiklikler nispeten keşfedilmemiş bir alan kalır.
Dinamik disülfür exchange ve thiol Kimya supramolecular yapıları yüzeyde yeni ve işlevsel Biyomalzeme verim potansiyeline sahip bir alandır. 1,2-dithiolane moieties (genellikle bir türevi lipoic asit (la) veya asparagusic asit (aa)) birleşme, lipozom sistemleri10,11, blok kopolimerler12,13ve olarak bildirilmiştir çapa yüzeyler14,15düzenleme. Burada, biz sentezi ve karakterizasyonu, N-terminus bir 1,2-dithiolane fonksiyonel grup16ile değiştirilir Alzheimer Hastaligi ile ilişkili Aβ peptid nucleating çekirdeğinden elde edilen bir kendi kendine montaj peptid raporu, 17. Elde edilen supramolecular lifleri şimdi amiloid lifleri18supramolecular yüzeyde disülfür-Satım ve thiol reaktivite çalışmaya deneysel bir platform olarak hizmet vermektedir.

<table>
	<tbody>
		<tr>
			<td>Rink amide MBHA resin, high load</td>
			<td>Gyros Protein Technologies</td>
			<td>RAM-5-HL</td>
			<td>Avoid contact with skin and eyes; do not inhale</td>
		</tr>
		<tr>
			<td>N,N-Dimethylformamide</td>
			<td>Fisher Scientific</td>
			<td>D119-4</td>
			<td>Flammable liquid and vapor; irritating to eyes and skin; Use personal protective equipment; keep away from open flame</td>
		</tr>
		<tr>
			<td>Fmoc-L-Val-OH</td>
			<td>Gyros Protein Technologies</td>
			<td>FLA-25-V</td>
			<td>Wear personal protective equipment; do not inhale</td>
		</tr>
		<tr>
			<td>Fmoc-L-Leu-OH</td>
			<td>Gyros Protein Technologies</td>
			<td>FLA-25-L</td>
			<td>Wear personal protective equipment; do not inhale</td>
		</tr>
		<tr>
			<td>Fmoc-L-Lys(Boc)-OH</td>
			<td>Gyros Protein Technologies</td>
			<td>FLA-25-KBC</td>
			<td>Wear personal protective equipment; do not inhale</td>
		</tr>
		<tr>
			<td>Fmoc-L-Phe-OH</td>
			<td>Gyros Protein Technologies</td>
			<td>FLA-25-F</td>
			<td>Wear personal protective equipment; do not inhale</td>
		</tr>
		<tr>
			<td>Fmoc-L-Ala-OH</td>
			<td>Gyros Protein Technologies</td>
			<td>FLA-25-A</td>
			<td>Wear personal protective equipment; do not inhale</td>
		</tr>
		<tr>
			<td>Fmoc-L-Gln(Trt)-OH</td>
			<td>Gyros Protein Technologies</td>
			<td>FLA-25-QT</td>
			<td>Wear personal protective equipment; do not inhale</td>
		</tr>
		<tr>
			<td>N,N,N&#8242;,N&#8242;-Tetramethyl-O-(1H-benzotriazol-1-yl)uronium hexafluorophosphate</td>
			<td>Gyros Protein Technologies</td>
			<td>26432</td>
			<td>Causes skin, eye and respiratory irritation; do not inhale; use under hood or in well ventilated area</td>
		</tr>
		<tr>
			<td>0.4 M N-methylmorpholine in DMF</td>
			<td>Gyros Protein Technologies</td>
			<td>PS3-MM-L</td>
			<td>highly flammable; wear personal protective equipment; keep away from heat and keep container tightly closed; do not inhale or swallow; wash skin thoroughly after handling</td>
		</tr>
		<tr>
			<td>20% piperidine in DMF</td>
			<td>Gyros Protein Technologies</td>
			<td>PS3-PPR-L</td>
			<td>Causes severe eye and skin burns; Flammable Liquid and vapor; Do not inhale</td>
		</tr>
		<tr>
			<td>dichloromethane</td>
			<td>Fisher Scientific</td>
			<td>D37-4</td>
			<td>May cause cancer; Do not inhale; Wear personal protective equipment; use under hood only; if contacted rise with water for at least 15 minutes and obtain medical attention</td>
		</tr>
		<tr>
			<td>acetonitrile</td>
			<td>Fisher Scientific</td>
			<td>A998-4</td>
			<td>Flammable; irritating to eyes; Use personal protective equipment; Use only under a fume hood; keep away from open flame or hot surface; if contacted rinse wiith water for at least 15 minutes and obtain medical attention</td>
		</tr>
		<tr>
			<td>trifluoroacetic acid</td>
			<td>Fisher Scientific</td>
			<td>A116-50</td>
			<td>Causes severe burns; do not inhale; harmful to aquatic life; use personal protective equipment; use only under fume hood; if contacted rinse with water for at least 15 minutes and obain immediate medical attention</td>
		</tr>
		<tr>
			<td>4% uranyl acetate</td>
			<td>Electron Microscopy Sciences</td>
			<td>22400-4</td>
			<td>Do not inhale; harmful to aquatic life</td>
		</tr>
		<tr>
			<td>4-(2-Hydroxyethyl)piperazine-1-ethanesulfonic acid</td>
			<td>Acros Organics</td>
			<td>AC172571000</td>
			<td>Do not inhale; use outdoors or in well-ventilated area</td>
		</tr>
		<tr>
			<td>nitrogen Gas</td>
			<td>TechAir</td>
			<td></td>
			<td>Contents under pressure, may explode if heated</td>
		</tr>
		<tr>
			<td>3-bromo-2-(bromomethyl)propionic acid</td>
			<td>Alfa Aesar</td>
			<td>AAA1963014</td>
			<td>Do not inhale; causes irritation to skin and eyes; corrosive</td>
		</tr>
		<tr>
			<td>sodium hydroxide</td>
			<td>Fisher Scientific</td>
			<td>S318-100</td>
			<td>Use personal protective equipment; use only under fume hood; if contact rinse area for at least 15 minutes and obtain medical attention</td>
		</tr>
		<tr>
			<td>potassium thioacetate</td>
			<td>Acros Organics</td>
			<td>AC221300250</td>
			<td>Causes skin and eye irritation; do not inhale; use personal protective equipment</td>
		</tr>
		<tr>
			<td>sulfuric acid</td>
			<td>Fisher Scientific</td>
			<td>SA213</td>
			<td>Causes burns; keep away from water; keep away from combustible material; do not inhale; use personal protective equipment; if contact rinse area for at least 15 minutes and obtain medical attention</td>
		</tr>
		<tr>
			<td>chloroform-d</td>
			<td>Acros Organics</td>
			<td>AC320690075</td>
			<td>Possible cancer hazard; irritating to skin and eyes; do not inhale; Use personal protective equipment; use only under fume hood; If contact rinse area for at least 15 minutes and obtain medical attention</td>
		</tr>
		<tr>
			<td>chloroform</td>
			<td>Fisher Scientific</td>
			<td>C298-4</td>
			<td>Possible cancer hazard; irritating to skin and eyes; do not inhale; Use personal protective equipment; use only under fume hood; If contact rinse area for at least 15 minutes and obtain medical attention</td>
		</tr>
		<tr>
			<td>N,N-diisopropylethylamine</td>
			<td>Acros Organics</td>
			<td>AC367841000</td>
			<td>Highly flammable; harmful to aquatic life; wear personal protective equipment; do not swallow</td>
		</tr>
		<tr>
			<td>ammonium hydroxide</td>
			<td>Fisher Scientific</td>
			<td>A669S-500</td>
			<td>Corrosive; do not inhale</td>
		</tr>
		<tr>
			<td>methanol</td>
			<td>Fisher Scientific</td>
			<td>A452-4</td>
			<td>Flammable liquid and vapor; use personal protective equipment; do not inhale; If contact rinse area for at least 15 minutes and obtain medical attention</td>
		</tr>
		<tr>
			<td>triisopropylsilane</td>
			<td>Sigma Aldrich</td>
			<td>233781</td>
			<td>Flammable; use personal proctective safety equipment; keep container tightly closed</td>
		</tr>
		<tr>
			<td>diethyl ether</td>
			<td>Fisher Scientific</td>
			<td>E138-1</td>
			<td>Extremely flammable; Irritating to skin and eyes; Use personal protective equipment</td>
		</tr>
		<tr>
			<td>2,5-dihydroxybenzoic acid</td>
			<td>Sigma Aldrich</td>
			<td>39319-10x10MG-F</td>
			<td>do not inhale; irritating to skin and eyes</td>
		</tr>
		<tr>
			<td>alpha-cyano-4-hydroxycinnamic acid</td>
			<td>Alfa Aesar</td>
			<td>AAJ67635EXK</td>
			<td></td>
		</tr>
		<tr>
			<td>c18 zip-tip</td>
			<td>Millipore</td>
			<td>ZTC18S096</td>
			<td></td>
		</tr>
		<tr>
			<td>tris(2-carboxyethyl) phospine hydrochloride</td>
			<td>Thermo Scientific</td>
			<td>PI20490</td>
			<td></td>
		</tr>
		<tr>
			<td>silica gel 60 F254 coated aluminum-backed TLC sheets</td>
			<td>EMD Millipore</td>
			<td>1.05549.0001</td>
			<td></td>
		</tr>
		<tr>
			<td>Thin walled Precision NMR tubes</td>
			<td>Bel-Art</td>
			<td>663000585</td>
			<td>5mm O.D.</td>
		</tr>
		<tr>
			<td>All-plastic Norm-Ject syringes</td>
			<td>Air Tite</td>
			<td>AL10</td>
			<td></td>
		</tr>
		<tr>
			<td>single-use needle</td>
			<td>BD PrecisionGlide</td>
			<td>BD 305185</td>
			<td>used needles get disposed on in sharps waste container</td>
		</tr>
		<tr>
			<td>disposable fritted syringe</td>
			<td>Torviq</td>
			<td>SF1000LL</td>
			<td>10mL fritted syringes were used in the report, but larger syringes are avaibale if needed for larger scale synthesis.</td>
		</tr>
		<tr>
			<td>carbon grid</td>
			<td>Ted Pella, Inc.</td>
			<td>CF200-CU</td>
			<td>Make sure to prepare samples and staining on the carbon grid side, not the shiny copper side of grid</td>
		</tr>
		<tr>
			<td>self-closing tweezers</td>
			<td>Electron Microscopy Sciences</td>
			<td>78318-3X</td>
			<td>very sharp tips, length: 120 mm</td>
		</tr>
		<tr>
			<td>0.1 mm short path length cell</td>
			<td>Starna Cells, Inc.</td>
			<td>20/C-Q-0.1</td>
			<td>Fragile</td>
		</tr>
		<tr>
			<td>10mL Vessel Caps</td>
			<td>CEM</td>
			<td>909210</td>
			<td></td>
		</tr>
		<tr>
			<td>10mL Pressure Vessels</td>
			<td>CEM</td>
			<td>908035</td>
			<td></td>
		</tr>
		<tr>
			<td>Aeris Semi-Prep HPLC column</td>
			<td>Phenomenex</td>
			<td>00F-4632-N0</td>
			<td>150 x 10mm</td>
		</tr>
		<tr>
			<td>cell holder</td>
			<td>Starna Cells, Inc.</td>
			<td>CH-2049</td>
			<td>Needed when using short pathlength cells</td>
		</tr>
		<tr>
			<td>PS3 peptide synthesizer</td>
			<td>Gyros Protein Technologies</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>DiscoverSP Microwave Reactor</td>
			<td>CEM</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>centrifuge</td>
			<td>HERMLE</td>
			<td>Z 206 A</td>
			<td>used a fixed 6x50 mL rotor</td>
		</tr>
		<tr>
			<td>HPLC</td>
			<td>Shimadzu</td>
			<td></td>
			<td>UV Detector</td>
		</tr>
		<tr>
			<td>nuclear magnetic resonance spectrometer</td>
			<td>Avance, Bruker</td>
			<td></td>
			<td>300 MHz</td>
		</tr>
		<tr>
			<td>MALDI-TOF mass spectrometer</td>
			<td>Axima Confidence, Shimadzu</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>lyophilizer</td>
			<td>Millrock Technology</td>
			<td>BT85A</td>
			<td></td>
		</tr>
		<tr>
			<td>Fourier-Transform Infrared Spectrometer</td>
			<td>Alpha Tensor, Bruker</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Transmission Electron Microscope</td>
			<td>Tecnai Spirit, FEI</td>
			<td></td>
			<td>Used with Gatan Orius Fiberoptic CCD digital camera. Accessed at CSCU Center for Nanotechnology</td>
		</tr>
		<tr>
			<td>Circular Dichroism Spectropolarimeter</td>
			<td>J-810, JASCO</td>
			<td></td>
			<td>Used with a six-cell Peltier temperature controller. Accessed at Wesleyan University.</td>
		</tr>
	</tbody>
</table>

synthesis and characterization of 1,2-dithiolane modified self-assembling peptides

Kendi kendine montaj peptid ve sonucu olan karakterizasyonu bir N-terminus 1,2-dithiolane sentezi üzerinde duruluyor değiştiren bu rapor supramolecular yapıları kendi kendine monte. Katı faz peptid sentez dithiolane habercisi molekülü 3-(acetylthio) -2-reçine MANŞONLA sentetik rota yararlanır-(acetylthiomethyl) propanoic asit ve mikrodalga destekli thioacetate deprotection peptid N-terminus reçine üzerinden son bölünme önce 1,2-dithiolane vermeye peptid değiştiren. Alzheimer hastalığı, ilişkili Aβ peptid nucleating çekirdeğinden elde edilen 1,2-dithiolane peptid, yüksek performanslı sıvı kromatografi (HPLC) arıtma sonra peptid çapraz-β amiloid lifleri kendi kendine montajı gösterilmiştir. Fourier dönüşümü kızılötesi spektroskopi (FT-IR), dairesel dichroism spektroskopisi (CD) ve transmisyon elektron mikroskobu (TEM) amiloid lifler karakterize etmek için protokolleri mevcuttur. N-terminal geliştirmeleriyle 1,2-dithiolane yan iyi karakterize kendi kendine montaj peptidler yöntemleri şimdi sonrası derleme değişiklik stratejiler geliştirmek ve dinamik kovalent kimya üzerinde keşfetmek için model sistemleri olarak keşfedilmeyi olabilir Supramolecular peptid nanofiber yüzeyler.

Dithiolane öncül molekül ilk tek adımlı sentezi kenara 1,2-dithiolane değiştiren peptid sentez geri kalanı sağlam destek (Şekil 1A) oluşur. 3 dönüşüm-bromo - 2-(bromomethyl) propiyonik asit 3-(acetylthio) -2-(acetylthiomethyl) propanoic asit, dithiolane habercisi 1tarafından H doğruladı ve 13C NMR (Şekil 1B ve C) daha önce ücretsiz için birleştiğinde Bir peptid hala-r...

Watch this Scientific Journal Video about Synthesis and Characterization of 1,2-Dithiolane Modified Self-Assembling Peptides at JoVE.com

Synthesis and Characterization of 1,2-Dithiolane Modified Self-Assembling Peptides

1,2-dithiolane sentezi için bir protokol peptid ve peptid kendinden montajlı kaynaklanan supramolecular yapıları karakterizasyonu değiştiren.

1,2-Dithiolane malzemelerin ve kendi kendine montaj peptidler değiştirilme tarihi

This report focuses on the synthesis of an N-terminus 1,2-dithiolane modified self-assembling peptide and the characterization of the resulting ...

synthesis-characterization-12-dithiolane-modified-self-assembling

Research

JoVE Journal

Chemistry

7.1K Görüntüleme.  Fairfield University. 1,2-dithiolane sentezi için bir protokol peptid ve peptid kendinden montajlı kaynaklanan supramolecular yapıları karakterizasyonu değiştiren.

Video: Synthesis and Characterization of 1,2-Dithiolane Modified Self-Assembling Peptides

Split-and-pool Synthesis and Characterization of Peptide Tertiary Amide Library

Peptidomimetics are great sources of protein ligands. The oligomeric nature of these compounds enables us to access large synthetic libraries on solid phase by using combinatorial chemistry. One of the most well studied classes of peptidomimetics is peptoids. Peptoids are easy to synthesize and have been shown to be proteolysis-resistant and cell-permeable. Over the past decade, many useful protein ligands have been identified through screening of peptoid libraries. However, most of the ligands identified from peptoid libraries do not display high affinity, with rare exceptions. This may be due, in part, to the lack of chiral centers and conformational constraints in peptoid molecules. Recently, we described a new synthetic route to access peptide tertiary amides (PTAs). PTAs are a superfamily of peptidomimetics that include but are not limited to peptides, peptoids and N-methylated peptides. With side chains on both &#945;-carbon and main chain nitrogen atoms, the conformation of these molecules are greatly constrained by sterical hindrance and allylic 1,3 strain. (Figure 1) Our study suggests that these PTA molecules are highly structured in solution and can be used to identify protein ligands. We believe that these molecules can be a future source of high-affinity protein ligands. Here we describe the synthetic method combining the power of both split-and-pool and sub-monomer strategies to synthesize a sample one-bead one-compound (OBOC) library of PTAs.

Peptide tertiary amides (PTAs) are a superfamily of peptidomimetics that include but are not limited to peptides, peptoids and N-methylated peptides. Here we describe a synthetic method which combines&#160;both split-and-pool and sub-monomer strategies to synthesize a one-bead one-compound library of PTAs.

Split-ve-havuz Sentezi ve Peptit Tersiyer Amid Kütüphanesi Karakterizasyonu

Peptidomimetics are great sources of protein ligands. The oligomeric nature of these compounds enables us to access large synthetic libraries on solid ...

Kimya

Nucleoside Triphosphates - From Synthesis to Biochemical Characterization

The traditional strategy for the introduction of chemical functionalities is the use of solid-phase synthesis by appending suitably modified phosphoramidite precursors to the nascent chain. However, the conditions used during the synthesis and the restriction to rather short sequences hamper the applicability of this methodology. On the other hand, modified nucleoside triphosphates are activated building blocks that have been employed for the mild introduction of numerous functional groups into nucleic acids, a strategy that paves the way for the use of modified nucleic acids in a wide-ranging palette of practical applications such as functional tagging and generation of ribozymes and DNAzymes. One of the major challenges resides in the intricacy of the methodology leading to the isolation and characterization of these nucleoside analogues.
	
	In this video article, we present a detailed protocol for the synthesis of these modified analogues using phosphorous(III)-based reagents. In addition, the procedure for their biochemical characterization is divulged, with a special emphasis on primer extension reactions and TdT tailing polymerization. This detailed protocol will be of use for the crafting of modified dNTPs and their further use in chemical biology.

The protocol described herein aims to explain and abridge the numerous obstacles in the way of the intricate route leading to modified nucleoside triphosphates. Consequently, this protocol facilitates both the synthesis of these activated building-blocks and their availability for practical applications.

Nükleosid trifosfatlar - sentezinden Biyokimyasal Karakterizasyonu için

The  traditional strategy for the introduction of chemical functionalities is the  use of solid-phase synthesis by appending suitably modified ...

Synthesis and Characterization of Functionalized Metal-organic Frameworks

Metal-organic frameworks have attracted extraordinary amounts of research attention, as they are attractive candidates for numerous industrial and technological applications. Their signature property is their ultrahigh porosity, which however imparts a series of challenges when it comes to both constructing them and working with them. Securing desired MOF chemical and physical functionality by linker/node assembly into a highly porous framework of choice can pose difficulties, as less porous and more thermodynamically stable congeners (e.g., other crystalline polymorphs, catenated analogues) are often preferentially obtained by conventional synthesis methods. Once the desired product is obtained, its characterization often requires specialized techniques that address complications potentially arising from, for example, guest-molecule loss or preferential orientation of microcrystallites. Finally, accessing the large voids inside the MOFs for use in applications that involve gases can be problematic, as frameworks may be subject to collapse during removal of solvent molecules (remnants of solvothermal synthesis). In this paper, we describe synthesis and characterization methods routinely utilized in our lab either to solve or circumvent these issues. The methods include solvent-assisted linker exchange, powder X-ray diffraction in capillaries, and materials activation (cavity evacuation) by supercritical CO2 drying. Finally, we provide a protocol for determining a suitable pressure region for applying the Brunauer-Emmett-Teller analysis to nitrogen isotherms, so as to estimate surface area of MOFs with good accuracy.

Synthesis, activation, and characterization of intentionally designed metal-organic framework materials is challenging, especially when building blocks are incompatible or unwanted polymorphs are thermodynamically favored over desired forms. We describe how applications of solvent-assisted linker exchange, powder X-ray diffraction in capillaries and activation via supercritical CO2 drying, can address some of these challenges.

Fonksiyonlu Sentezi ve Karakterizasyonu Metal-organik Altyapıları

Metal-organic frameworks have attracted extraordinary amounts of research attention, as they are attractive candidates for numerous industrial and ...

The Synthesis, Characterization and Reactivity of a Series of Ruthenium N-triphosPh Complexes

Herein we report the synthesis of a tridentate phosphine ligand N(CH2PPh2)3 (N-triphosPh) (1) via a phosphorus based Mannich reaction of the hydroxylmethylene phosphine precursor with ammonia in methanol under a nitrogen atmosphere. The N-triphosPh ligand precipitates from the solution after approximately 1 hr of reflux and can be isolated analytically pure via simple cannula filtration procedure under nitrogen. Reaction of the N-triphosPh ligand with [Ru3(CO)12] under reflux affords a deep red solution that show evolution of CO gas on ligand complexation. Orange crystals of the complex [Ru(CO)2{N(CH2PPh2)3}-&#954;3P] (2) were isolated on cooling to RT. The 31P{1H} NMR spectrum showed a characteristic single peak at lower frequency compared to the free ligand. Reaction of a toluene solution of complex 2 with oxygen resulted in the instantaneous precipitation of the carbonate complex [Ru(CO3)(CO){N(CH2PPh2)3}-&#954;3P] (3) as an air stable orange solid. Subsequent hydrogenation of 3 under 15 bar of hydrogen in a high-pressure reactor gave the dihydride complex [RuH2(CO){N(CH2PPh2)3}-&#954;3P] (4), which was fully characterized by X-ray crystallography and NMR spectroscopy. Complexes 3 and 4 are potentially useful catalyst precursors for a range of hydrogenation reactions, including biomass-derived products such as levulinic acid (LA). Complex 4 was found to cleanly react with LA in the presence of the proton source additive NH4PF6 to give [Ru(CO){N(CH2PPh2)3}-&#954;3P{CH3CO(CH2)2CO2H}-&#954;2O](PF6) (6).

The Synthesis, Characterization and Reactivity of a Series of Ruthenium N-triphosPh Complexes

Ruthenium phosphine complexes are widely used for homogeneous catalytic reactions such as hydrogenations. The synthesis of a series of novel tridentate ruthenium complexes bearing the N-triphos ligand N(CH2PPh2)3 is reported. Additionally, the stoichiometric reaction of a dihydride Ru&#8211;N-triphos complex with levulinic acid is described.

Rutenyum bir Serisi Sentezi, Karakterizasyonu ve Tepkime<em&gt; K</em&gt; -triphos<sup&gt; Ph</sup&gt; Kompleksi

Herein we report the synthesis of a tridentate phosphine ligand N(CH2PPh2)3 (N-triphosPh) (1) via a phosphorus based Mannich reaction of the ...

Synthesis, Characterization, and Functionalization of Hybrid Au/CdS and Au/ZnS Core/Shell Nanoparticles

Plasmonic nanoparticles are an attractive material for light harvesting applications due to their easily modified surface, high surface area and large extinction coefficients which can be tuned across the visible spectrum. Research into the plasmonic enhancement of optical transitions has become popular, due to the possibility of altering and in some cases improving photo-absorption or emission properties of nearby chromophores such as molecular dyes or quantum dots. The electric field of the plasmon can couple with the excitation dipole of a chromophore, perturbing the electronic states involved in the transition and leading to increased absorption and emission rates. These enhancements can also be negated at close distances by energy transfer mechanism, making the spatial arrangement of the two species critical. Ultimately, enhancement of light harvesting efficiency in plasmonic solar cells could lead to thinner and, therefore, lower cost devices. The development of hybrid core/shell particles could offer a solution to this issue. The addition of a dielectric spacer between a gold nanoparticles and a chromophore is the proposed method to control the exciton plasmon coupling strength and thereby balance losses with the plasmonic gains. A detailed procedure for the coating of gold nanoparticles with CdS and ZnS semiconductor shells is presented. The nanoparticles show high uniformity with size control in both the core gold particles and shell species allowing for a more accurate investigation into the plasmonic enhancement of external chromophores.

The synthesis of uniform gold nanoparticles coated with semiconductor shells of CdS or ZnS is performed. The semiconductor coating is conducted by first depositing a silver sulfide shell and exchanging the silver cations for zinc or cadmium cations.

Sentezi, Karakterizasyonu ve Hibrid Au / CdS ve Au / ZnS Çekirdek / Shell Nanopartiküller işlevselleştirilmesi

Plasmonic nanoparticles are an attractive material for light harvesting applications due to their easily modified surface, high surface area and large ...

Synthesis and Characterization of Supramolecular Colloids

Control over colloidal assembly is of utmost importance for the development of functional colloidal materials with tailored structural and mechanical properties for applications in photonics, drug delivery and coating technology. Here we present a new family of colloidal building blocks, coined supramolecular colloids, whose self-assembly is controlled through surface-functionalization with a benzene-1,3,5-tricarboxamide (BTA) derived supramolecular moiety. Such BTAs interact via directional, strong, yet reversible hydrogen-bonds with other identical BTAs. Herein, a protocol is presented that describes how to couple these BTAs to colloids and how to quantify the number of coupling sites, which determines the multivalency of the supramolecular colloids. Light scattering measurements show that the refractive index of the colloids is almost matched with that of the solvent, which strongly reduces the van der Waals forces between the colloids. Before photo-activation, the colloids remain well dispersed, as the BTAs are equipped with a photo-labile group that blocks the formation of hydrogen-bonds. Controlled deprotection with UV-light activates the short-range hydrogen-bonds between the BTAs, which triggers the colloidal self-assembly. The evolution from the dispersed state to the clustered state is monitored by confocal microscopy. These results are further quantified by image analysis with simple routines using ImageJ and Matlab. This merger of supramolecular chemistry and colloidal science offers a direct route towards light- and thermo-responsive colloidal assembly encoded in the surface-grafted monolayer.

A protocol for the synthesis and characterization of colloids coated with supramolecular moieties is described. These supramolecular colloids undergo self-assembly upon the activation of the hydrogen-bonds between the surface-anchored molecules by UV-light.

Supramoleküler Kolloidlerin Sentezi ve Karakterizasyonu

Control over colloidal assembly is of utmost importance for the development of functional colloidal materials with tailored structural and mechanical ...

Synthesis and Characterization of Fe-doped Aluminosilicate Nanotubes with Enhanced Electron Conductive Properties

The goal of the protocol is to synthesize Fe-doped aluminosilicate nanotubes of the imogolite type with the formula (OH)3Al2-xFexO3SiOH. Doping with Fe aims at lowering the band gap of imogolite, an insulator with the chemical formula (OH)3Al2O3SiOH, and at modifying its adsorption properties towards azo-dyes, an important class of organic pollutants of both wastewater and groundwater.
Fe-doped nanotubes are obtained in two ways: by direct synthesis, where FeCl3 is added to an aqueous mixture of the Si and Al precursors, and by post-synthesis loading, where preformed nanotubes are put in contact with a FeCl3&#8226;6H2O aqueous solution. In both synthesis methods, isomorphic substitution of Al3+ by Fe3+ occurs, preserving the nanotube structure. Isomorphic substitution is indeed limited to a mass fraction of ~1.0% Fe, since at a higher Fe content (i.e., a mass fraction of 1.4% Fe), Fe2O3 clusters form, especially when the loading procedure is adopted. The physicochemical properties of the materials are studied by means of X-ray powder diffraction (XRD), N2 sorption isotherms at -196 &#176;C, high resolution transmission electron microscopy (HRTEM), diffuse reflectance (DR) UV-Vis spectroscopy, and &#950;-potential measurements. The most relevant result is the possibility to replace Al3+ ions (located on the outer surface of the nanotubes) by post-synthesis loading on preformed imogolite without perturbing the delicate hydrolysis equilibria occurring during nanotube formation. During the loading procedure, an anionic exchange occurs, where Al3+ ions on the outer surface of the nanotubes are replaced by Fe3+ ions. In Fe-doped aluminosilicate nanotubes, isomorphic substitution of Al3+ by Fe3+ is found to affect the band gap of doped imogolite. Nonetheless, Fe3+ sites on the outer surface of nanotubes are able to coordinate organic moieties, like the azo-dye Acid Orange 7, through a ligand-displacement mechanism occurring in an aqueous solution.

Here, we present a protocol to synthesize and characterize Fe-doped aluminosilicate nanotubes. The materials are obtained by either sol-gel synthesis upon addition of FeCl3&#8226;6H2O to the mixture containing the Si and Al precursors or by post-synthesis ionic exchange of preformed aluminosilicate nanotubes.

Geliştirilmiş Elektron İletken özellikleri ile Fe-katkılı Alüminosilikat Nanotüpler Sentezi ve Karakterizasyonu

The goal of the protocol is to synthesize Fe-doped aluminosilicate nanotubes of the imogolite type with the formula (OH)3Al2-xFexO3SiOH. Doping with Fe ...

Self-assembling Morphologies Obtained from Helical Polycarbodiimide Copolymers and Their Triazole Derivatives

A facile method for the preparation of polycarbodiimide-based secondary structures (e.g., nano-rings, &#34;craters,&#34; fibers, looped fibers, fibrous networks, ribbons, worm-like aggregates, toroidal structures, and spherical particles) is described. These aggregates are morphologically influenced by extensive hydrophobic side chain-side chain interactions of the singular polycarbodiimide strands, as inferred by atomic force microscopy (AFM) and scanning electron microscopy (SEM) techniques. Polycarbodiimide-g-polystyrene copolymers (PS-PCDs) were prepared by a combination of synthetic methods, including coordination-insertion polymerization, copper(I)-catalyzed azide alkyne cycloaddition (CuAAC) &#34;click&#34; chemistry, and atom transfer radical polymerization (ATRP). PS-PCDs were found to form specific toroidal architectures at low concentrations in CHCl3. To determine the influence of a more polar solvent medium (i.e., THF and THF/EtOH) on polymer aggregation behavior, a number of representative PS-PCD composites have been tested to show discrete concentration-dependent spherical particles. These fundamental studies are of practical interest to the development of experimental procedures for desirable architectures by directed self-assembly in thin film. These architectures may be exploited as drug carriers, whereas other morphological findings represent certain interest in the area of novel functional materials.

Here, we present a protocol to prepare and visualize secondary structures (e.g., fibers, toroidal architectures, and nano-spheres) derived from helical polycarbodiimides. The morphology characterized by both atomic force microscopy (AFM) and scanning electron microscopy (SEM) was shown to depend on molecular structure, concentration, and the solvent of choice.

Sarmal polikarbodiimidden Kopolimer ve Bunların Triazole Türevleri elde edilen kendinden montaj morfolojileri

A facile method for the preparation of polycarbodiimide-based secondary structures (e.g., nano-rings, &#34;craters,&#34; fibers, looped fibers, fibrous ...

Chemical Precipitation Method for the Synthesis of Nb2O5 Modified Bulk Nickel Catalysts with High Specific Surface Area

We demonstrate a method for the synthesis of NixNb1-xO catalysts with sponge-like and fold-like nanostructures. By varying the Nb:Ni ratio, a series of NixNb1-xO nanoparticles with different atomic compositions (x = 0.03, 0.08, 0.15, and 0.20) have been prepared by chemical precipitation. These NixNb1-xO catalysts are characterized by X-ray diffraction, X-ray photoelectron spectroscopy, and scanning electron microscopy. The study revealed the sponge-like and fold-like appearance of Ni0.97Nb0.03O and Ni0.92Nb0.08O on the NiO surface, and the larger surface area of these NixNb1-xO catalysts, compared with the bulk NiO. Maximum surface area of 173 m2/g can be obtained for Ni0.92Nb0.08O catalysts. In addition, the catalytic hydroconversion of lignin-derived compounds using the synthesized Ni0.92Nb0.08O catalysts have been investigated.

Chemical Precipitation Method for the Synthesis of Nb2O5 Modified Bulk Nickel Catalysts with High Specific Surface Area

A protocol for the synthesis of sponge-like and fold-like Ni1-xNbxO nanoparticles by chemical precipitation is presented.

Nb2O5 sentezi için kimyasal yağış yöntemi toplu nikel katalizör yüksek belirli yüzey alanı ile modifiye

We demonstrate a method for the synthesis of NixNb1-xO catalysts with sponge-like and fold-like nanostructures. By varying the Nb:Ni ratio, a series of ...

Facile Protocol for the Synthesis of Self-assembling Polyamine-based Peptide Amphiphiles (PPAs) and Related Biomaterials

Polyamine-based Peptide Amphiphiles (PPAs) are a new class of self-assembling amphiphilic biomaterials-related to the peptide amphiphiles (PAs). Traditional PAs possess charged amino acids as solubilizing groups (lysine, arginine), which are directly connected to a lipid segment or can contain a linker region made of neutral amino acids. Tuning the peptide sequence of PAs can yield diverse morphologies. Similarly, PPAs possess a hydrophobic segment and neutral amino acids, but also contain polyamine molecules as water solubilizing (hydrophilic) groups. As is the case with PAs, PPAs can also self-assemble into diverse morphologies, including small rods, twisted nano-ribbons, and fused nano-sheets, when dissolved in water. However, the presence of both primary and secondary amines on a single polyamine molecule poses a significant challenge when synthesizing PPAs. In this paper, we show a simple protocol, based on literature precedents, to achieve a facile synthesis of PPAs using solid phase peptide synthesis (SPPS). This protocol can be extended to the synthesis of PAs and other similar systems. We also illustrate the steps that are needed for cleavage from the resin, identification, and purification.

Facile Protocol for the Synthesis of Self-assembling Polyamine-based Peptide Amphiphiles PPAs and Related Biomaterials

The synthesis of polyamine-based peptide amphiphiles (PPAs) is a significant challenge due to the presence of multiple amine nitrogens, which requires judicious use of protecting groups to mask these reactive functionalities. In this paper, we describe a facile method for the preparation of these new class of self-assembling molecules.

1,2-Dithiolane malzemelerin ve kendi kendine montaj peptidler değiştirilme tarihi

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