TY - JOUR
T1 - Apparent growth phase-dependent phosphorylation of malonyl coenzyme A:acyl carrier protein transacylase (MCAT), a major fatty acid synthase II component in Mycobacterium bovis BCG
AU - Sinha, Indrajit
AU - Boon, Calvin
AU - Dick, Thomas
N1 - Funding Information: We thank Robert Qi, Proteomics Laboratory, IMCB, for mass spectrometric identification of MCAT, and Alice Tay, Genome Analysis Laboratory, IMCB, for DNA sequencing. We are grateful to Robin J. Philp, Bioprocessing Technology Centre, Singapore, for his help and suggestions. We thank Westley Kee for help in the initial phase of the project. We would like to thank B. Gicquel for pJEM15. T.D. is an adjunct staff member of the Department of Microbiology, National University of Singapore. This work was supported by the Agency for Science, Technology and Research (A*STAR), Singapore.
PY - 2003/10/10
Y1 - 2003/10/10
N2 - Probing protein extracts from exponentially growing and stationary phase cultures of Mycobacterium bovis BCG with anti-phospho amino acid antibodies revealed a 31-kDa anti-phospho threonine antibody-reactive protein specific to growing culture. The corresponding protein was purified via two-dimensional gel electrophoresis and identified via mass spectrometry to be malonyl coenzyme A:acyl carrier protein transacylase (MCAT), a component of the fatty acid biosynthetic pathway. MCAT tagged with histidine reacted with anti-phospho threonine antibody and was positive in an in-gel chemical assay for phospho proteins. Analysis of the growth phase dependence of MCAT-His phosphorylation and protein levels showed that phosphorylated MCAT-His can be detected only in growing culture. In contrast, MCAT-His protein level was growth phase-independent. These results suggest that MCAT may be a substrate of a protein kinase and phosphatase, and that aspects of fatty acid synthesis in tubercle bacilli are regulated by protein phosphorylation.
AB - Probing protein extracts from exponentially growing and stationary phase cultures of Mycobacterium bovis BCG with anti-phospho amino acid antibodies revealed a 31-kDa anti-phospho threonine antibody-reactive protein specific to growing culture. The corresponding protein was purified via two-dimensional gel electrophoresis and identified via mass spectrometry to be malonyl coenzyme A:acyl carrier protein transacylase (MCAT), a component of the fatty acid biosynthetic pathway. MCAT tagged with histidine reacted with anti-phospho threonine antibody and was positive in an in-gel chemical assay for phospho proteins. Analysis of the growth phase dependence of MCAT-His phosphorylation and protein levels showed that phosphorylated MCAT-His can be detected only in growing culture. In contrast, MCAT-His protein level was growth phase-independent. These results suggest that MCAT may be a substrate of a protein kinase and phosphatase, and that aspects of fatty acid synthesis in tubercle bacilli are regulated by protein phosphorylation.
KW - Fatty acid synthesis
KW - Mycobacterium
KW - Protein phosphorylation
UR - https://www.scopus.com/pages/publications/1542648496
UR - https://www.scopus.com/pages/publications/1542648496#tab=citedBy
U2 - 10.1016/S0378-1097(03)00664-5
DO - 10.1016/S0378-1097(03)00664-5
M3 - Article
C2 - 14568160
SN - 0378-1097
VL - 227
SP - 141
EP - 147
JO - FEMS Microbiology Letters
JF - FEMS Microbiology Letters
IS - 1
ER -