下载PDF
{"title":"Quantitative Analysis of Surface Expression of Membrane Proteins Using Cold-Adapted Proteases","authors":"Faraz Ahmad, Kai Kaila, Peter Blaesse","doi":"10.1002/0471140864.ps0311s73","DOIUrl":null,"url":null,"abstract":"<p>This unit presents an improved method for quantitative analysis of surface expression of membrane proteins utilizing a cold-adapted trypsin. Preservation of the proteolytic activity of the enzyme at 0° to 4°C allows cleavage of surface-expressed membrane proteins at temperatures at which trafficking of the mammalian plasmalemmal proteins is blocked. This provides an important advantage over established trypsin-cleavage protocols since it can be applied to membrane proteins with a fast turnover rate of the membrane pool and a fast recycling rate. Compared to surface biotinylation, the method is less time consuming. <i>Curr. Protoc. Protein Sci</i>. 73:3.11.1-3.11.12. © 2013 by JohnWiley & Sons, Inc.</p>","PeriodicalId":10866,"journal":{"name":"Current Protocols in Protein Science","volume":"73 1","pages":""},"PeriodicalIF":0.0000,"publicationDate":"2018-02-16","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1002/0471140864.ps0311s73","citationCount":"0","resultStr":null,"platform":"Semanticscholar","paperid":null,"PeriodicalName":"Current Protocols in Protein Science","FirstCategoryId":"1085","ListUrlMain":"https://onlinelibrary.wiley.com/doi/10.1002/0471140864.ps0311s73","RegionNum":0,"RegionCategory":null,"ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"Q1","JCRName":"Biochemistry, Genetics and Molecular Biology","Score":null,"Total":0}
引用次数: 0
引用
批量引用
Abstract
This unit presents an improved method for quantitative analysis of surface expression of membrane proteins utilizing a cold-adapted trypsin. Preservation of the proteolytic activity of the enzyme at 0° to 4°C allows cleavage of surface-expressed membrane proteins at temperatures at which trafficking of the mammalian plasmalemmal proteins is blocked. This provides an important advantage over established trypsin-cleavage protocols since it can be applied to membrane proteins with a fast turnover rate of the membrane pool and a fast recycling rate. Compared to surface biotinylation, the method is less time consuming. Curr. Protoc. Protein Sci . 73:3.11.1-3.11.12. © 2013 by JohnWiley & Sons, Inc.
利用冷适应蛋白酶定量分析膜蛋白的表面表达
本单元提出了一种改进的方法，利用冷适应胰蛋白酶对膜蛋白的表面表达进行定量分析。在0°至4°C下保持酶的蛋白水解活性，可以在阻断哺乳动物浆哺乳动物蛋白运输的温度下切割表面表达的膜蛋白。这提供了一个重要的优势，超过已建立的胰蛋白酶切割方案，因为它可以应用于膜蛋白具有快速的膜池周转率和快速的再循环率。与表面生物素化相比，该方法耗时更少。咕咕叫。Protoc。蛋白质科学。73:3.11.1-3.11.12。©2013 by JohnWiley &儿子,Inc。
本文章由计算机程序翻译，如有差异，请以英文原文为准。
京公网安备 11010802042870号
京ICP备2023020795号-1
分享
求助内容：
应助结果提醒方式：
扫码关注我们
