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CFP10 discriminates between nonacetylated and acetylated ESAT-6 of Mycobacterium tuberculosis by differential interaction

Item Type:Article
Title:CFP10 discriminates between nonacetylated and acetylated ESAT-6 of Mycobacterium tuberculosis by differential interaction
Creators Name:Okkels, L.M. and Mueller, E.C. and Schmid, M. and Rosenkrands, I. and Kaufmann, S.H.E. and Andersen, P. and Jungblut, P.R.
Abstract:ESAT-6 (the 6 kDa early secreted antigenic target) protein species in short-term culture filtrate of Mycobacterium tuberculosis were separated in a 4-5 narrow range p/ gradient two-dimensional gel electrophoresis (2-DE). Eight ESAT-6 protein species were analyzed in detail by peptide mass fingerprinting matrix-assisted laser desorption/ionization-mass spectrometry as well as by electrospray ionization-tandem mass spectrometry. An N-terminal Thr acetylation was identified in four species and a C-terminal truncation was identified in two species. In 2-DE blot overlay assays, the recombinant 10 kDa culture filtrate protein (CFP10) discriminated N-terminal acetylated and nonacetylated ESAT-6 by differential interaction, whereas removal of the C-terminal 11 residues of ESAT-6 had no effects thereon. This example shows that the access to the protein species level can be a prerequisite to understand regulation of protein-protein interaction.
Keywords:Narrow Range PH Gradient, Post-Translational Modification, Two-Dimensional Blot Overlay
Source:Proteomics
ISSN:1615-9853
Publisher:Wiley (Germany)
Volume:4
Number:10
Page Range:2954-2960
Date:1 January 2004
Official Publication:https://doi.org/10.1002/pmic.200400906
PubMed:View item in PubMed

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