Name: interactions with and membrane permeabilization of brain mitochondria by amyloid fibrils
Uploaded: 2019-09-28T14:00:00
Description: Watch this Scientific Journal Video about Interactions with and Membrane Permeabilization of Brain Mitochondria by Amyloid Fibrils at JoVE.com

这项工作得到了伊朗赞詹基础科学高级研究所研究理事会的资助。

所有动物实验均按照德黑兰大学医学研究所动物护理和使用委员会（IACUC）进行。通过锐化断头刀刃和采用果断和迅速的叶片运动，尽最大努力尽量减少对大鼠的痛苦和有害影响。
1. 大脑均质化和线粒体隔离
注：所有线粒体隔离试剂均根据西姆斯和安德森14号制备。
<ol><li>为线粒体隔离准备缓冲液<ol>
<li>准备 100 mM Tris-HCl 溶液：重量为 ...

<ol>
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F., Morero, R. D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Lysozyme+interactions+with+phospholipid+vesicles:+relationships+with+fusion+and+release+of+aqueous+content.">Lysozyme interactions with phospholipid vesicles: relationships with fusion and release of aqueous content.</a> Biochimica et Biophysica Acta. 1193, 101-106 (1994).</li><li>Roqanian, S., 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=Polyphenols+protect+mitochondrial+membrane+against+permeabilization+induced+by+HEWL+oligomers:+possible+mechanism+of+action.">Polyphenols protect mitochondrial membrane against permeabilization induced by HEWL oligomers: possible mechanism of action.</a> International Journal of Biological Macromolecules. 103, 709-720 (2017).</li><li>Ulmer, T. S., Bax, A., Cole, N. B., Nussbaum, R. L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Structure+and+dynamics+of+micelle-bound+human+alphasynuclein.">Structure and dynamics of micelle-bound human alphasynuclein.</a> The Journal of Biological Chemistry. 280, 9595-9603 (2005).</li><li>Stockl, M., Fischer, P., Wanker, E., Herrmann, A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Alpha-synuclein+selectively+binds+to+anionic+phospholipids+embedded+in+liquid-disordered+domains.">Alpha-synuclein selectively binds to anionic phospholipids embedded in liquid-disordered domains.</a> Journal of Molecular Biology. 375, 1394-1404 (2008).</li><li>Devi, L., 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=Mitochondrial+import+and+accumulation+of+&#945;-synuclein+impair+complex+I+in+human+dopaminergic+neuronal+cultures+and+Parkinson+disease+brain.">Mitochondrial import and accumulation of &#945;-synuclein impair complex I in human dopaminergic neuronal cultures and Parkinson disease brain.</a> The Journal of Biological Chemistry. 283, 9089-9100 (2008).</li><li>Ghio, S., Kamp, F., Cauchi, R., Giese, A., Vassallo, N. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Interaction+of+&#945;-synuclein+with+biomembranes+in+Parkinson's+disease-role+of+cardiolipin.">Interaction of &#945;-synuclein with biomembranes in Parkinson's disease-role of cardiolipin.</a> Progress in Lipid Research. 61, 73-82 (2016).</li><li>Petersen, C. A. H., 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=The+amyloid+&#946;-peptide+is+imported+into+mitochondria+via+the+TOM+import+machinery+and+localized+to+mitochondrial+cristae.">The amyloid &#946;-peptide is imported into mitochondria via the TOM import machinery and localized to mitochondrial cristae.</a> Proceedings of National Academy of Sciences of the United States of America. 105, 13145-13150 (2008).</li><li>Costa, V., Scorrano, L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Shaping+the+role+of+mitochondria+in+the+pathogenesis+of+Huntington's+disease.">Shaping the role of mitochondria in the pathogenesis of Huntington's disease.</a> EMBO Journal. 31, 1853-1864 (2012).</li><li>Vande Velde, C., Miller, T. M., Cashman, N. R., Cleveland, D. 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大量的实验结果支持了纤维细胞聚集体细胞毒性与其与生物膜4、5相互作用和渗透的能力显著相关的假设。然而，大多数数据都基于人工脂质双层，不一定反映生物膜的内在特性，生物膜是具有多种磷脂和蛋白质的异构结构。本文以脑线粒体作为体外生物膜，介绍了一个在膜水平上研究纤维集体细胞毒性的模型。
在实验中，重要的?...

淀粉样蛋白相关疾病，称为淀粉样蛋白，构成由不同组织和器官中不溶性蛋白质沉积的出现所定义的一大群疾病1，2。其中，神经退行性疾病是蛋白质聚集出现在中枢或周围神经系统2中最常见的形式。虽然已经提出一些机制参与淀粉样粒体的毒性3，越来越多的证据表明细胞膜中断和渗透作为淀粉样蛋白病理学的主要机制4， 5.除血浆膜外，内部细胞器（即线粒体）也可能受到影响。
有趣的是，新的证据表明线粒体功能障碍在神经退行性疾病的发病机制中起着关键作用，包括阿尔茨海默氏症和帕金森病6，7。根据这个问题，许多报告表明淀粉样蛋白β-肽、β-核糖核酸、亨廷顿蛋白和ALS相关突变SOD1蛋白与线粒体8、9、10的结合和积累。 11.淀粉样粒体渗透膜的机制被认为是通过离散通道（孔）的形成和/或通过非特异性洗涤剂状机制5、12、 13.值得注意的是，这些结论大多基于涉及磷脂模型系统的报告，而直接针对生物膜中发生的事件的研究很少见。显然，这些人造脂质双层不一定反映生物膜的内在特性，包括线粒体，线粒体是异质结构，由各种各样的磷脂和蛋白质组成。
在本研究中，从大鼠大脑中分离的线粒体被用作体外生物模型，以检查β-核糖核酸（作为淀粉样蛋白）、牛胰岛素（作为模型肽显示的淀粉样纤维素）的破坏性影响。与注射局部淀粉样蛋白病有关的人类胰岛素的重要结构同源性，以及母鸡蛋白液化酶（HEWL;作为研究淀粉样蛋白聚集的常见模型蛋白）。然后通过观察线粒体麻化脱氢酶（MDH）（位于线粒体基质）和线粒体活性氧的释放，研究淀粉样纤维诱导的线粒体膜的相互作用和可能的损害物种（ROS）增强。

<table>
	<tbody>
		<tr>
			<td>2&#8242;,7&#8242;-Dichlorodihydrofluorescein diacetate</td>
			<td>Sigma</td>
			<td>35845</td>
			<td></td>
		</tr>
		<tr>
			<td>Ammonium sulfate</td>
			<td>Merck</td>
			<td>1012171000</td>
			<td></td>
		</tr>
		<tr>
			<td>Black 96-well plate</td>
			<td>Corning</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Black Clear-bottomed 96-well plate</td>
			<td>Corning</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Bovine insulin</td>
			<td>Sigma</td>
			<td>I6634</td>
			<td></td>
		</tr>
		<tr>
			<td>Bovine Serum Albumin (BSA)</td>
			<td>Sigma</td>
			<td>A2153</td>
			<td></td>
		</tr>
		<tr>
			<td>BSA essentially fatty acid-free</td>
			<td>Sigma</td>
			<td>A6003</td>
			<td></td>
		</tr>
		<tr>
			<td>Centrifuge</td>
			<td>Sigma</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Crystal clear sealing tape</td>
			<td>Corning</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>CuSO4</td>
			<td>Sigma</td>
			<td>451657</td>
			<td></td>
		</tr>
		<tr>
			<td>Dialysis bag (cut off 2 KDa)</td>
			<td>Sigma</td>
			<td>D2272</td>
			<td></td>
		</tr>
		<tr>
			<td>Dounce homogenizer</td>
			<td>Potter Elvehjem</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>EDTA</td>
			<td>Sigma</td>
			<td>E9884</td>
			<td></td>
		</tr>
		<tr>
			<td>Fluorescence plate reader</td>
			<td>BioTek</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Fluorescence spectrophotometer</td>
			<td>Cary Eclipse VARIAN</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Folin</td>
			<td>Merck</td>
			<td>F9252</td>
			<td></td>
		</tr>
		<tr>
			<td>Glycine</td>
			<td>Sigma</td>
			<td>G7126</td>
			<td></td>
		</tr>
		<tr>
			<td>Guillotine</td>
			<td>Made in Iran</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>HCl</td>
			<td>Merck</td>
			<td>H1758</td>
			<td></td>
		</tr>
		<tr>
			<td>Hen Egg White Lysozyme (HEWL)</td>
			<td>Sigma</td>
			<td>L6876</td>
			<td></td>
		</tr>
		<tr>
			<td>Na2CO3</td>
			<td>Sigma</td>
			<td>S7795</td>
			<td></td>
		</tr>
		<tr>
			<td>NaH2PO4</td>
			<td>Sigma</td>
			<td>S7907</td>
			<td></td>
		</tr>
		<tr>
			<td>NaOH</td>
			<td>Merck</td>
			<td>S8045</td>
			<td></td>
		</tr>
		<tr>
			<td>Oxaloacetate</td>
			<td>Sigma</td>
			<td>O4126</td>
			<td></td>
		</tr>
		<tr>
			<td>Percoll</td>
			<td>GE Healthcare</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Phosphate Buffer Saline (PBS)</td>
			<td>Sigma</td>
			<td>CS0030</td>
			<td></td>
		</tr>
		<tr>
			<td>PMSF</td>
			<td>Sigma</td>
			<td>P7626</td>
			<td></td>
		</tr>
		<tr>
			<td>Potassium sodium tartrate</td>
			<td>Sigma</td>
			<td>217255</td>
			<td></td>
		</tr>
		<tr>
			<td>Quartz cuvette</td>
			<td>Sigma</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Spectrophotometer</td>
			<td>analytik jena</td>
			<td>SPEKOL 2000 model</td>
			<td></td>
		</tr>
		<tr>
			<td>Succinate</td>
			<td>Sigma</td>
			<td>S2378</td>
			<td></td>
		</tr>
		<tr>
			<td>Sucrose</td>
			<td>Merck</td>
			<td>1076871000</td>
			<td></td>
		</tr>
		<tr>
			<td>Thermomixer</td>
			<td>Eppendorph</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Thioflavin T</td>
			<td>Sigma</td>
			<td>T3516</td>
			<td></td>
		</tr>
		<tr>
			<td>Tris-HCl</td>
			<td>Merck</td>
			<td>1082191000</td>
			<td></td>
		</tr>
		<tr>
			<td>Triton X-100</td>
			<td>Sigma</td>
			<td>T9284</td>
			<td></td>
		</tr>
		<tr>
			<td>Tryptone</td>
			<td>QUELAB</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Water bath</td>
			<td>Memmert</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Yeast Extract</td>
			<td>QUELAB</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>&#946;-NADH</td>
			<td>Sigma</td>
			<td>N8129</td>
			<td></td>
		</tr>
	</tbody>
</table>

interactions with and membrane permeabilization of brain mitochondria by amyloid fibrils

越来越多的证据表明，膜渗透，包括线粒体等内膜，是神经退行性疾病淀粉样体性引起的毒性的常见特征和主要机制。然而，大多数描述膜中断机制的报告都基于磷脂模型系统，而直接针对生物膜水平发生的事件的研究很少见。本文介绍了一个模型，用于研究膜层淀粉样蛋白毒性机制。对于线粒体隔离，密度梯度培养基用于获得以最小的骨髓污染获得制剂。线粒体膜完整性确认后，研究β-核糖核酸、牛胰岛素和母鸡蛋白溶解酶（HEWL）与大鼠脑线粒体（作为体外生物模型）产生的淀粉样纤维蛋白的相互作用。结果表明，用纤维束体处理脑线粒体可引起不同程度的膜渗透和ROS含量增强。这表明淀粉样纤维蛋白和线粒体膜之间的结构相关相互作用。建议淀粉样纤维的生物物理特性及其与线粒体膜的具体结合，可以解释其中一些观测结果。

该协议描述了一个模型，用于研究淀粉样纤维蛋白与大鼠脑线粒体作为体外生物模型的相互作用。对于线粒体制剂，15%（v/v）密度梯度培养基用于去除骨髓，作为脑组织的主要污染14。如图1A所示，在30，700 x g处离心产生了两个不同的物质带，即骨髓（作为波段1的主要成分）和带2，它们含有丰富的线粒体分数。
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Watch this Scientific Journal Video about Interactions with and Membrane Permeabilization of Brain Mitochondria by Amyloid Fibrils at JoVE.com

Interactions with and Membrane Permeabilization of Brain Mitochondria by Amyloid Fibrils

这里提供了一个协议，用于研究从不同组织和大脑不同区域分离的来自不同组织和不同区域的线粒体的不同肽和蛋白质的原生形态、前纤维和成熟的淀粉样纤维之间的相互作用。

淀粉样蛋白纤维蛋白的相互作用和脑线粒体的膜渗透

A growing body of evidence indicates that membrane permeabilization, including internal membranes such as mitochondria, is a common feature and primary ...

在这段视频中，我们描述了一个模型研究，利用大鼠脑线粒体作为体外生物模型，在膜水平上淀粉样毒性的机理。尽管累积报告描述了淀粉样蛋白聚合在磷脂模型系统中的膜扰动机制，但直接针对从开发生物膜开始的事件进行了研究。在目前的视频中，我们描述α-核素的淀粉样纤维分析大鼠脑线粒体膜作为体外生物模型。我们认为，线粒体作为具有良好特征的膜的细胞器，可以为与神经退行性疾病相关的膜水平上淀粉样细胞毒性机制的分子研究提供极其有用的生物模型系统。用小动物断头台斩首大鼠，并在斩首后一分钟内将大脑从规模上除名，以限制线粒体特性的可能恶化。重要的是要快速工作，并保持一切冰上在整个程序。用30毫升的隔离缓冲液清洗组织两次。转移到含有冷隔离缓冲液的烧杯中，用剪刀细细切碎大脑。将组织悬浮液转移到20毫升冷均匀均质器。使用九次上下冲程与电动害虫使组织碎片均匀。将混合物留在冰上约30秒后，每组三个均质冲程，以确保均质保持寒冷。将均质转移到预冷却的离心管中，在四摄氏度时以1，300 G的离心机进行三分钟。小心地将上一液式液 dec，然后转移到预冷却的离心管中。在4摄氏度时以21，000G离心10分钟。丢弃上流剂，用移液器轻轻搅拌混合物，在密度梯度介质中重新填充颗粒。在4摄氏度的固定角度转子中，在30，700G下离心5分钟。使用巴斯德移液器，去除顶部积聚的尖锐末端，其中大多含有米林。在线粒体分数中加入八毫升隔离缓冲液，同时用移液器轻轻搅拌混合物。在 4 摄氏度时在 16，700 G 下离心 10 分钟。小心地拆下上清液，使底部松动的颗粒不受干扰。将每毫升10毫克无脂肪酸BSA加入离心管，同时用移液器的尖端轻轻搅拌混合物。通过添加隔离缓冲区将体积调整为 5 毫升。在 4 摄氏度时以 6，900 G 离心 10 分钟。这应该会产生一个坚定的颗粒。去干上经，轻轻在隔离缓冲液中重新填充线粒。用冷隔离缓冲液稀释线粒体同质质至每毫升1毫克，并放在两个1.5毫升管中，通常每管195微升线粒体。将五微升20%体积每体积Triton X-100添加到一个管中，作为最大酶活性的正对照，将5微升去压水添加到另一个管中进行控制，然后与搅拌器混合。在设置为 30 摄氏度的温水浴中孵育管子 10 分钟。在16000G四摄氏度的固定角度转子中，通过固定角转子的管离心来降粒线粒体15分钟。使用下一部分描述的标准分光光度测定法，仔细收集产生的超生物质，以评估线粒体麦芽酸脱氢酶的活性。计算线粒体膜的完整性如下。用于麦芽酸脱氢酶活性测定，移液器890和880微升50毫摩尔三叶草-HCL缓冲液分别对空白和样品测试库维特，其次是100微升50毫摩尔氧化乙酸酯和10微升10毫摩尔β-NADH。将铜光计放入分光光度计中，并参考空白。加入10微升线粒体同质每毫升1毫克的样品。立即通过反转混合。在 340 纳米下，NADH 氧化导致的吸光率下降记录一分钟。对于α-核素淀粉样颤动，添加294微升的蛋白质溶液，200微摩尔到每1.5毫升管。然后添加六微升 ThT 溶液一毫摩尔。在800 RPM的恒搅拌下，在37摄氏度的热混合器中混合溶液并孵育管。对于牛胰岛素淀粉样纤维颤动，加入637微升的蛋白质溶液，250微摩尔到1.5毫升管。然后加入13微升ThT溶液一毫摩尔，然后搅拌。在透明底部 96 井板的每个井中加入 200 微升的蛋白质溶液。用水晶般清晰的密封胶带密封板。将板装入 Cyation 5 荧光板读卡器。以 30 分钟间隔测量 ThT 荧光 12 小时。在440纳米时使用激发，在485纳米时使用发射。选择井。每次测量前摇动板五秒钟。将温度设置为 57 摄氏度，无搅拌。准备两个系列1.5毫升管含有线粒体同质酸盐，一个系列用于MDH释放测定，另一个用于线粒体ROS测量。将α-核素、牛胰岛素或母鸡蛋清裂酶的新鲜或淀粉样纤维加入线粒体同质质，然后轻轻移液。在温水浴中，含有线粒体悬浮液的孵育管30分钟，设置为30摄氏度。对于麦芽酸脱氢酶释放测定，离心机在16，000G下孵育线粒体异质体15分钟。仔细收集产生的超级钠，以测定线粒体MDH的活性，如协议第四节所述。计算 MDH 的释放，如下所示。对于线粒体ROS测量，移液器191微升孵育线粒体均质到96井板的每个井。添加四微升 50 微摩尔 DCFDA。然后添加5微升200毫摩尔成功。在温水浴中孵育板30分钟，设置为30摄氏度，同时轻轻搅拌。将板装入 Cyation 5 荧光板读卡器，根据协议部分测量荧光强度。通过测量三元 X-100 在膜中断和酶释放之前和之后，通过测量分离线粒体的 MDH 活性，评估线粒体膜完整性。如你所看到的，我们通常发现线粒体制剂是大约93%的完好无损。进行了荧光测定，以监测淀粉样纤维的生长。三种蛋白质的淀粉样蛋白颤动曲线显示了典型的西格莫德模式，分别到达约96、12和96小时的高原，分别用于α-核素、牛胰岛素和母鸡蛋清裂酶，这表明淀粉样蛋白纤维的形成得到了AFM测量的更证实。通过线粒体MDH和线粒体ROS测量的释放，对线粒体纤维对线粒体膜的可能渗透和损伤进行了调查。正如你在左图中看到的，在线粒体中加入α-核素淀粉样纤维后观察到MDH的实质性释放。虽然胰岛素纤维检测到轻微的释放，但母鸡蛋清裂酶纤维被发现无效。右图显示淀粉样纤维对线粒体ROS含量的影响。虽然在添加母鸡蛋清糖酶或胰岛素淀粉样纤维纤维后，线粒体ROS没有显著增强，但使用α-核素纤维素治疗导致脑线粒体的ROS含量显著增加。本视频介绍了膜水平纤维聚集细胞毒性机制的模型研究，演示了由各种蛋白质产生的淀粉样纤维如何引起不同程度的膜损伤和渗透。虽然淀粉样纤维及其特定的结合膜的一些主题似乎提供了与膜相互作用的能力，并导致破坏稳定和随后的扰动。在这段视频之后，您将能够研究不同肽的原生形式、纤维前和成熟淀粉样纤维的相互作用，以及从不同组织或大脑不同区域分离的线粒体。

Research

JoVE Journal

Biochemistry

Using Scaffold Liposomes to Reconstitute Lipid-proximal Protein-protein Interactions In Vitro

Using Scaffold Liposomes to Reconstitute Lipid-proximal Protein-protein Interactions In Vitro

使用脚手架脂质体重构脂质近端蛋白质 - 蛋白质相互作用<em&gt;体外</em

Studies of integral membrane proteins in vitro are frequently complicated by the presence of a hydrophobic transmembrane domain. Further complicating ...

科研

生物化学

Method to Visualize and Analyze Membrane Interacting Proteins by Transmission Electron Microscopy

方法可视化和用透射电子显微镜分析膜蛋白的相互作用

Monotopic proteins exert their function when attached to a membrane surface, and such interactions depend on the specific lipid composition and on the ...

A Tailored HPLC Purification Protocol That Yields High-purity Amyloid Beta 42 and Amyloid Beta 40 Peptides, Capable of Oligomer Formation

一个定制HPLC纯化协议，产量高纯度淀粉样蛋白β42和贝塔淀粉样蛋白肽40，有能力寡聚物形成的

Amyloidogenic peptides such as the Alzheimer's disease-implicated Amyloid beta (A&#946;), can present a significant challenge when trying to obtain high ...

Imaging Amyloid Tissues Stained with Luminescent Conjugated Oligothiophenes by Hyperspectral Confocal Microscopy and Fluorescence Lifetime Imaging

用高光谱共聚焦显微镜和荧光寿命成像对发光共轭吩染色的淀粉样组织的成像

Proteins that deposit as amyloid in tissues throughout the body can be the cause or consequence of a large number of diseases. Among these we find ...

Isolation and Respiratory Measurements of Mitochondria from Arabidopsis thaliana

Isolation and Respiratory Measurements of Mitochondria from Arabidopsis thaliana

拟南芥线粒体的分离和呼吸测量

Mitochondria are essential organelles involved in numerous metabolic pathways in plants, most notably the production of adenosine triphosphate (ATP) from ...

Rab10 Phosphorylation Detection by LRRK2 Activity Using SDS-PAGE with a Phosphate-binding Tag

用磷酸盐结合标记的 SDS-页 Rab10 磷酸化检测 LRRK2 活性

Mutations in leucine-rich repeat kinase 2 (LRRK2) have been shown to be linked with familial Parkinson&#39;s disease (FPD). Since abnormal activation of ...

Detection of Detergent-sensitive Interactions Between Membrane Proteins

膜蛋白对洗涤剂敏感相互作用的检测

Our ability to explore protein-protein interactions is the key to understanding regulatory connections in the cell. However, detection of protein-protein ...

Use of Two Dimensional Semi-denaturing Detergent Agarose Gel Electrophoresis to Confirm Size Heterogeneity of Amyloid or Amyloid-like Fibers

二维半变性洗涤剂琼脂糖凝胶电泳法确定淀粉样或淀粉样纤维的粒度异质性

Amyloid or amyloid-like fibers have been associated with many human diseases, and are now being discovered to be important for many signaling pathways. ...

Generation of Native, Untagged Huntingtin Exon1 Monomer and Fibrils Using a SUMO Fusion Strategy

利用相扑融合策略生成国产、未加标签的杭丁顿蛋白 Exon1 单体和纤维

Huntington&#39;s Disease (HD) is an inherited fatal neurodegenerative disease caused by a CAG expansion (&#8805;36) in the first exon of the HD gene, ...

Detection of Small GTPase Prenylation and GTP Binding Using Membrane Fractionation and  GTPase-linked Immunosorbent Assay

膜分馏和 gtpase-关联免疫吸附法检测小 gtpase 预基化和 gtp 结合

The Rho GTPase family belongs to the Ras superfamily and includes approximately 20 members in humans. Rho GTPases are important in the regulation of ...