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CFP10 discriminates between nonacetylated and acetylated ESAT‐6 of Mycobacterium tuberculosis by differential interaction
Author(s) -
Okkels Limei Meng,
Müller EvaChristina,
Schmid Monika,
Rosenkrands Ida,
Kaufmann Stefan H. E.,
Andersen Peter,
Jungblut Peter R.
Publication year - 2004
Publication title -
proteomics
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.26
H-Index - 167
eISSN - 1615-9861
pISSN - 1615-9853
DOI - 10.1002/pmic.200400906
Subject(s) - esat 6 , mycobacterium tuberculosis , acetylation , mass spectrometry , electrospray ionization , peptide , chemistry , tandem mass spectrometry , peptide mass fingerprinting , western blot , recombinant dna , microbiology and biotechnology , biology , chromatography , biochemistry , tuberculosis , proteomics , medicine , pathology , gene
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 I 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.