Efficient Protein Digestion at Elevated Temperature in the Presence of Sodium Dodecyl Sulfate and Calcium Ions for Membrane Proteomics

被引：4

作者：

Loraine, Jessica ^{[1
]}

Alhumaidan, Ohoud ^{[1
,2
,3
]}

Bottrill, Andrew R. ^{[4
,5
]}

Mistry, Sharad C. ^{[4
]}

Andrew, Peter ^{[1
,2
]}

Mukamolova, Galina V. ^{[1
,2
]}

Turapov, Obolbek ^{[1
,2
]}

机构：

[1] Univ Leicester, Dept Infect Immun & Inflammat, Leicester LE1 9HN, Leics, England

[2] Univ Leicester, Dept Resp Sci, Leicester LE1 9HN, Leics, England

[3] King Saud Univ, Dept Clin Lab Sci, Riyadh 11433, Saudi Arabia

[4] Univ Leicester, Prot Nucle Acid Chem Lab, Leicester LE1 7RH, Leics, England

[5] Univ Warwick, Sch Life Sci, Prote Res Technol Platform, Coventry CV4 7AL, W Midlands, England

来源：

ANALYTICAL CHEMISTRY | 2019年 / 91卷 / 15期

基金：

英国生物技术与生命科学研究理事会;

关键词：

MASS-SPECTROMETRY; STATISTICAL-MODEL; TANDEM; MS/MS;

D O I：

10.1021/acs.analchem.9b00484

中图分类号：

O65 [分析化学];

学科分类号：

070302 ; 081704 ;

摘要：

The growing significance of membrane proteins inspires continuous development and improvement of methods for robust membrane proteomics. Here, we developed a very simple and efficient method for membrane protein digestion using an ionic detergent, sodium dodecyl sulfate (SDS), at high temperature, conditions where trypsin is normally inactivated. Our results suggest that trypsin can be stabilized by a combination of calcium ions and sodium chloride, which enables protein digestion at elevated temperature in the presence of strong ionic detergents such as SDS. Finding the conditions for stabilization of trypsin offers novel opportunities for the application of detergents for the investigation of membrane proteins.

引用

页码：9516 / 9521

页数：6

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