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Isolation of a 43 kDa protein from Mycobacterium tuberculosis H 37 Rv and its identification as a pyridine nucleotide transhydrogenase
Author(s) -
Deshpande R.G.,
Khan M.B.,
Bhat D.A.,
Navalkar R.G.
Publication year - 1994
Publication title -
journal of applied bacteriology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.889
H-Index - 156
eISSN - 1365-2672
pISSN - 0021-8847
DOI - 10.1111/j.1365-2672.1994.tb02813.x
Subject(s) - microbiology and biotechnology , mycobacterium bovis , biochemistry , mycobacterium tuberculosis , alanine , biology , escherichia coli , protein subunit , affinity chromatography , monoclonal antibody , peptide sequence , nucleic acid sequence , amino acid , enzyme , chemistry , antibody , tuberculosis , dna , gene , medicine , pathology , immunology
R.G. DESHPANDE, M.B. KHAN, D.A. BHAT AND R.G. NAVALKAR. 1994. A 43 kDa protein (TB43) was isolated from the cell sonicate (CS) of Mycobacterium tuberculosis H 37 Rv with immobilized metal affinity chromatography (IMAC) on a Ni‐nitrilotriacetic acid column. Two‐dimensional electrophoresis of the IMAC fraction showed a major spot with an M r of 43000 and a pI of ˜6.0. The N‐terminal amino acid sequence of TB43 was met‐arg‐val‐gly‐ile‐pro‐asn‐glu‐thr‐lys‐asn‐asn‐glu‐phe‐arg‐val‐ala‐ile‐thr‐pro‐ala. It showed 86% homology with the N‐terminal end of the alanine dehydrogenase of Myco. tuberculosis and 65% homology with the N‐terminal end of the alpha‐subunit of the Escherichia coli pyridine nucleotide transhydrogenase (Tsh). TB43 did not show any alanine dehydrogenase activity and did not react with monoclonal antibody (MAb) HBT10, which is known to recognize the 40 kDa alanine dehydrogenase of Myco. tuberculosis. It was also not recognized by MAb F29–29 which is known to react with a 43 kDa protein of Myco. tuberculosis complex. This protein exhibited strong Tsh activity. A similar 43 kDa protein showing Tsh activity was also isolated by IMAC from Myco. bovis CS. However, the pI of the protein was ˜7.0. A similar protein could not be isolated from the CS or culture filtrate of Myco. bovis BCG and Myco. tuberculosis H 37 Ra. TB43 is a cell‐associated pyridine nucleotide transhydrogenase and is distinct from the 40/44 kDa secreted alanine dehydrogenase of Myco. tuberculosis.