An improved method to unravel phosphoacceptors in Ser/Thr protein kinase-phosphorylated substrates.

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.

journal_name

Proteomics

journal_title

Proteomics

authors

Molle V,Leiba J,Zanella-Cléon I,Becchi M,Kremer L

doi

10.1002/pmic.201000316

subject

Has Abstract

pub_date

2010-11-01 00:00:00

pages

3910-5

issue

21

eissn

1615-9853

issn

1615-9861

journal_volume

10

pub_type

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