Name: split-ubiquitin based membrane yeast two-hybrid (myth) system: a powerful tool for identifying protein-protein interactions
Uploaded: 2010-02-01T16:25:00
Description: Watch this Scientific Journal Video about Split-Ubiquitin Based Membrane Yeast Two-Hybrid MYTH System: A Powerful Tool For Identifying Protein-Protein Interactions at JoVE.com

Biz Şafak Edmonds Bu yazının eleştirel bir okuma için teşekkür etmek istiyorum. Stagljar laboratuvar Kanada Yenilik (CFI), Kanada Sağlık Araştırmaları (CIHR) Enstitüsü, Kalp ve İnme Vakfı, Kanada Kanser Derneği ve Novartis Vakfı fonları tarafından desteklenmektedir.

1. Temel Bilgiler Protein-protein etkileşimleri (PPI) tüm hücresel süreçleri yöneten temel yapı taşlarıdır. Sonuç olarak, tüm etkileşimleri sıkıca bu biyolojik denge içinde bir değişim yaygın hastalığı ve kanser hücresi dönüşümün önemli bir rol oynar gibi, hücresel homeostazı korumak amacıyla düzenlenmiş olması esastır. Karmaşık sinyalleşme kaskadlar başlatmak ve arabuluculuk ile ilgili problemleri gibi sağlık alanında son önemi olmuştur ilaçlar da...

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	<li>Stagljar, I., Fields, S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Analysis+of+membrane+protein+interactions+using+yeast-based+technologies.">Analysis of membrane protein interactions using yeast-based technologies.</a> Trends Biochem Sci. 27 (11), 559-563 (2002).</li><li>Iyer, K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Utilizing+the+split-ubiquitin+membrane+yeast+two-hybrid+system+to+identify+protein-protein+interactions+of+integral+membrane+proteins.">Utilizing the split-ubiquitin membrane yeast two-hybrid system to identify protein-protein interactions of integral membrane proteins.</a> Sci STKE. 275, pl3-pl3 (2005).</li><li>Paumi, C. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Mapping+protein-protein+interactions+for+the+yeast+ABC+transporter+Ycf1p+by+integrated+split-ubiquitin+membrane+yeast+two-hybrid+analysis.">Mapping protein-protein interactions for the yeast ABC transporter Ycf1p by integrated split-ubiquitin membrane yeast two-hybrid analysis.</a> Mol Cell. 26 (1), 15-25 (2007).</li><li>Stagljar, I. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+genetic+system+based+on+split-ubiquitin+for+the+analysis+of+interactions+between+membrane+proteins+in+vivo.">A genetic system based on split-ubiquitin for the analysis of interactions between membrane proteins in vivo.</a> Proc Natl Acad Sci U S A. 95 (9), 5187-5192 (1998).</li><li>Gisler, S. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Monitoring+protein-protein+interactions+between+the+mammalian+integral+membrane+transporters+and+PDZ-interacting+partners+using+a+modified+split-ubiquitin+membrane+yeast+two-hybrid+system.">Monitoring protein-protein interactions between the mammalian integral membrane transporters and PDZ-interacting partners using a modified split-ubiquitin membrane yeast two-hybrid system.</a> Mol Cell Proteomics. 7 (7), 1362-1377 (2008).</li><li>Scheper, W. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Coordination+of+N-glycosylation+and+protein+translocation+across+the+endoplasmic+reticulum+membrane+by+Sss1+protein.">Coordination of N-glycosylation and protein translocation across the endoplasmic reticulum membrane by Sss1 protein.</a> J Biol Chem. 278 (39), 37998-38003 (2003).</li><li>Thaminy, S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Identification+of+novel+ErbB3-interacting+factors+using+the+split-ubiquitin+membrane+yeast+two-hybrid+system.">Identification of novel ErbB3-interacting factors using the split-ubiquitin membrane yeast two-hybrid system.</a> Genome Res. 13 (7), 1744-1753 (2003).</li><li>Johnsson, N., Varshavsky, A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Split+ubiquitin+as+a+sensor+of+protein+interactions+in+vivo.">Split ubiquitin as a sensor of protein interactions in vivo.</a> Proc Natl Acad Sci U S A. 91 (22), 10340-10344 (1994).</li><li>Kelleher, D. J., Gilmore, R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+Saccharomyces+cerevisiae+oligosaccharyltransferase+is+a+protein+complex+composed+of+Wbp1p,+Swp1p,+and+four+additional+polypeptides.">The Saccharomyces cerevisiae oligosaccharyltransferase is a protein complex composed of Wbp1p, Swp1p, and four additional polypeptides.</a> J Biol Chem. 269 (17), 12908-12917 (1994).</li><li>Chevallier, M. R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Cloning+and+transcriptional+control+of+a+eucaryotic+permease+gene.">Cloning and transcriptional control of a eucaryotic permease gene.</a> Mol Cell Biol. 2 (8), 977-984 (1982).</li></ol>

MİT, tam uzunlukta membran proteinleri ve sitozolik veya membrana bağlı ortaklar arasındaki etkileşimlerin belirlenmesi sağlayan ilk yüksek kapasiteli bir sistemdir. Membran proteini organizmalar [3-7] bir dizi çalışma için kullanılır olmuştur. Ancak, protein-faiz MİT ile çalışmak için uygun olduğundan emin olmak için incelenip gerekebilir belirli ayrıntıları vardır. Birçok membrana bağlı proteinler daha sonra olgun bir protein üretmek için bölünmüş bir siny...

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split-ubiquitin based membrane yeast two-hybrid (myth) system: a powerful tool for identifying protein-protein interactions

Integral membran proteinlerinin temel biyolojik ve klinik önemi tam uzunlukta transmembran proteinler için protein-protein etkileşimleri (ÜFE), yüksek verimli bir tanımlama için bir maya-tabanlı sistem geliştirme istenir. Bu amaçla, bizim laboratuvar split-ubikuitin Membran Maya İki Hybrid (MİT) sistemi geliştirdi. Bu teknoloji kullanarak geçici ve istikrarlı bir protein etkileşimleri hassas algılama sağlar<em&gt; Saccharomyces cerevisiae</em&gt; Konak organizma olarak. MİT ubikuitin iki istikrarlı moieties ayrılmış olabilir gözlem yararlanır: (maya C-terminal yarı ubikuitin C<sub&gt; Ub</sub&gt;) Ve N-terminal ubikuitin kısmın yarısı (N<sub&gt; Ub</sub&gt;). MİT, bu prensip protein-protein etkileşimleri &#39;sensör&#39; olarak kullanılmak üzere uyarlanmıştır. Kısaca, integral membran yem protein C eritilir.<sub&gt; Ub</sub&gt; Yapay bir transkripsiyon faktörü bağlıdır. Prey proteinler, ya bireysel ya da kütüphane biçimi N kaynaşmıştır<sub&gt; Ub</sub&gt; Benzer parçaları. Yem ve av arasındaki Protein etkileşimi, tam uzunlukta &#39;pseudo-ubikuitin&#39; molekülü oluşturan ubikuitin moieties sulandırıldıktan yol açar. Bu molekül, transkripsiyon faktörü ve raportör gen ekspresyonu sonraki indüksiyon bölünme sonucu, deubiquitinating sitozolik enzimler tarafından tanınan sırayla. Sistem son derece uyarlanabilir ve özellikle yüksek verimli tarama için çok uygundur. Bu maya ve diğer organizmalar hem de integral membran proteinleri kullanarak etkileşimleri araştırmak için başarılı bir işe alındı.

Watch this Scientific Journal Video about Split-Ubiquitin Based Membrane Yeast Two-Hybrid MYTH System: A Powerful Tool For Identifying Protein-Protein Interactions at JoVE.com

Split-Ubiquitin Based Membrane Yeast Two-Hybrid MYTH System: A Powerful Tool For Identifying Protein-Protein Interactions

MİT, geçici ve istikrarlı model organizma Saccharomyces cerevisiae ifade proteinler arasındaki etkileşimleri hassas algılama sağlar. Bu, yüksek kapasiteli bir şekilde etkileşim ortakları belirlemek amacıyla eksojen ve maya integral membran proteinleri çalışma başarıyla tatbik edilmiştir.

Split-Ubiquitin Dayalı Membran Maya İki Hybrid (MİT) Sistemi: protein-protein etkileşimleri belirlenmesi için güçlü bir araç

The fundamental biological and clinical importance of integral membrane proteins prompted the development of a yeast-based system for the high-throughput ...

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Split-Ubiquitin Based Membrane Yeast Two-Hybrid (MYTH) System: A Powerful Tool For Identifying Protein-Protein Interactions

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Progranulin (PGRN), also known as granulin epithelin precursor (GEP), is a 593-amino-acid autocrine growth factor. PGRN is known to play a critical role ...

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A Protocol for the Identification of Protein-protein Interactions Based on 15N Metabolic Labeling, Immunoprecipitation, Quantitative Mass Spectrometry and Affinity Modulation

Dayanarak protein-protein etkileşimleri ve kimliği için bir protokol<sup&gt; 15</sup&gt; K Metabolik Etiketleme, immünopresipitasyon, Kantitatif Kütle Spektrometrisi ve Affinity Modülasyon

Protein-protein interactions are fundamental for many biological processes in the cell. Therefore, their characterization plays an important role in ...

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Identifying Protein-protein Interaction in Drosophila Adult Heads by Tandem Affinity Purification TAP

Protein-protein etkileşimi belirlenmesi<em&gt; Drosophila</emTandem Affinity Arıtma tarafından&gt; Adult Başlar (TAP)

Genetic screens conducted using Drosophila melanogaster (fruit fly) have made numerous milestone discoveries in the advance of biological sciences. ...

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As a critical regulator of drug metabolism and inflammation, Pregnane X Receptor (PXR), plays an important role in disease pathophysiology linking ...

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Yakalama Bileşik Kütle Spektrometre - Roman c-di-GMP Efektör Proteinler tanımlamak için güçlü bir araç

Considerable progress has been made during the last decade towards the identification and characterization of enzymes involved in the synthesis ...

Investigating Protein-protein Interactions in Live Cells Using Bioluminescence Resonance Energy Transfer

Biyoparlaklık Rezonans Enerji Transferi kullanma Canlı Hücrelerde Protein-protein etkileşimleri araştıran

Assays based on Bioluminescence Resonance Energy Transfer (BRET) provide a sensitive and reliable means to monitor protein-protein interactions in live ...

Probing High-density Functional Protein Microarrays to Detect Protein-protein Interactions

Yüksek yoğunluklu Fonksiyonel Protein mikrodizileri Probing Protein-protein Etkileşimleri Algılama

High-density functional protein microarrays containing ~4,200 recombinant yeast proteins are examined for kinase protein-protein interactions using an ...

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Protein-protein interactions mediate most of the processes in the living cell and control homeostasis of the organism. Impaired protein interactions may ...