Item Type: | Article |
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Title: | CFP10 discriminates between nonacetylated and acetylated ESAT-6 of Mycobacterium tuberculosis by differential interaction |
Creators Name: | Okkels, L.M., Mueller, E.C., Schmid, M., Rosenkrands, I., Kaufmann, S.H.E., 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 |
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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