Affiliation:
1. Wuhan National Laboratory for Optoelectronics Huazhong University of Science and Technology Wuhan P. R. China
2. Key Laboratory of Magnetic Resonance in Biological System State Key Laboratory of Magnetic Resonance and Atomic and Molecular Physics National Center for Magnetic Resonance in Wuhan Innovation Academy for Precision Measurement Science and Technology Chinese Academy of Sciences Wuhan P. R. China
Abstract
AbstractOligomerization is an important feature of proteins, which gives a defined quaternary structure to complete the biological functions. Although frequently observed in membrane proteins, characterizing the oligomerization state remains complicated and time‐consuming. In this study, 0.05% (w/v) sarkosyl‐polyacrylamide gel electrophoresis (05SAR‐PAGE) was used to identify the oligomer states of the membrane proteins CpxA, EnvZ, and Ma‐Mscl with high sensitivity. Furthermore, two‐dimensional electrophoresis (05SAR/sodium dodecyl sulfate–PAGE) combined with western blotting and liquid chromatography–tandem mass spectrometry was successfully applied to study the complex of CpxA/OmpA in cell lysate. The results indicated that 05SAR‐PAGE is an efficient, economical, and practical gel method that can be widely used for the identification of membrane protein oligomerization and the analysis of weak protein interactions.
Funder
National Natural Science Foundation of China