An improved method to unravel phosphoacceptors in Ser/Thr protein kinase-phosphorylated substrates |
| |
Authors: | Molle Virginie Leiba Jade Zanella-Cléon Isabelle Becchi Michel Kremer Laurent |
| |
Institution: | Institut de Biologie et Chimie des Protéines (UMR), CNRS, Université Lyon1, BioSciences, Lyon-Gerland, Lyon, France. vmolle@ibcp.fr |
| |
Abstract: | Identification of the phosphorylated residues of bacterial Ser/Thr protein kinase (STPK) substrates still represents a challenging task. Herein, we present a new strategy allowing the rapid determination of phosphoacceptors in kinase substrates, essentially based on the dual expression of the kinase with its substrate in the surrogate E. coli, followed by MS analysis in a single-step procedure. The performance of this strategy is illustrated using two distinct proteins from Mycobacterium tuberculosis as model substrates, the GroEL2 and HspX chaperones. A comparative analysis with a standard method that includes mass spectrometry analysis of in vitro phosphorylated substrates is also addressed. |
| |
Keywords: | Mass spectrometry Microbiology Mycobacterium Phosphoproteomics Ser/Thr protein kinase |
本文献已被 PubMed 等数据库收录! |
|