title: Phosphorylation of Mycobacterium tuberculosis protein tyrosine kinase A PtkA by Ser/Thr protein kinases.
abstract: Mycobacterium tuberculosis (Mtb), the causative agent of tuberculosis (TB), has inflicted about one third of mankind and claims millions of deaths worldwide annually. Signalling plays an important role in Mtb pathogenesis and persistence, and thus represents attractive resource for drug target candidates. Here, we show that protein tyrosine kinase A (PtkA) can be phosphorylated by Mtb endogenous eukaryotic-like Ser/Thr protein kinases (eSTPKs). Kinase assays showed that PknA, PknD, PknF, and PknK can phosphorylate PtkA in dose- and time-dependent manner. Enzyme kinetics suggests that PknA has the highest affinity and enzymatic efficiency towards PtkA. Furthermore, protein-protein interaction assay in surrogate host showed that PtkA interacts with multi-eSTPKs in vivo, including PknA. Lastly, we show that PtkA phosphorylation by eSTPKs occurs on threonine residues and may effect tyrosine phosphorylation levels and thus PtkA activity in vitro. These results demonstrate that PtkA can serve as a substrate to many eSTPKs and suggests that's its activity can be regulated.
authors.affiliation: Department of Medicine, Division of Infectious Diseases, University of British Columbia, Vancouver, BC, V5Z 3J5, Canada; Institute of Modern Biopharmaceuticals, State Key Laboratory Breeding Base of Eco-Environment and Bio-Resource of the Three Gorges Area, School of Life Sciences, Southwest University, Chongqing, 400715, China; Institute of Ethnic-minority Medicine, School of Chemistry & Environmental Science, Guizhou Minzu University, Guiyang, 550025, China.
authors.forename: Peifu
authors.initials: P
authors.lastname: Zhou
authors.affiliation: Department of Medicine, Division of Infectious Diseases, University of British Columbia, Vancouver, BC, V5Z 3J5, Canada.
authors.forename: Dennis
authors.initials: D
authors.lastname: Wong
authors.affiliation: Department of Medicine, Division of Infectious Diseases, University of British Columbia, Vancouver, BC, V5Z 3J5, Canada; Institute of Modern Biopharmaceuticals, State Key Laboratory Breeding Base of Eco-Environment and Bio-Resource of the Three Gorges Area, School of Life Sciences, Southwest University, Chongqing, 400715, China.
authors.forename: Wu
authors.initials: W
authors.lastname: Li
authors.affiliation: Institute of Modern Biopharmaceuticals, State Key Laboratory Breeding Base of Eco-Environment and Bio-Resource of the Three Gorges Area, School of Life Sciences, Southwest University, Chongqing, 400715, China. Electronic address: georgex@swu.edu.cn.
authors.forename: Jianping
authors.initials: J
authors.lastname: Xie
authors.affiliation: Department of Medicine, Division of Infectious Diseases, University of British Columbia, Vancouver, BC, V5Z 3J5, Canada. Electronic address: yossi@mail.ubc.ca.
authors.forename: Yossef
authors.initials: Y
authors.lastname: Av-Gay
mesh_terms: D001426:Bacterial Proteins / Q000737:chemistry* / Q000378:metabolism; D015964:Gene Expression Regulation, Bacterial; D007527:Isoenzymes / Q000737:chemistry / Q000378:metabolism; D007700:Kinetics; D009169:Mycobacterium tuberculosis / Q000737:chemistry* / Q000201:enzymology; D010766:Phosphorylation; D011494:Protein Kinases / Q000737:chemistry / Q000378:metabolism; D017346:Protein Serine-Threonine Kinases / Q000737:chemistry* / Q000378:metabolism; D011505:Protein-Tyrosine Kinases / Q000737:chemistry* / Q000378:metabolism; D011994:Recombinant Proteins / Q000737:chemistry / Q000378:metabolism; D015398:Signal Transduction; D013912:Threonine / Q000737:chemistry / Q000378:metabolism
chemical_list: D001426:Bacterial Proteins; D007527:Isoenzymes; D011994:Recombinant Proteins; D013912:Threonine; D011494:Protein Kinases; C451020:PknA protein, Mycobacterium tuberculosis; C112455:PknD protein, Nostoc sp. PCC 7120; D011505:Protein-Tyrosine Kinases; C487814:PknF protein, Mycobacterium tuberculosis; C554363:Pknk protein, Mycobacterium tuberculosis; D017346:Protein Serine-Threonine Kinases
keywords: Eukaryotic-like Ser/Thr protein kinase; Mycobacterium tuberculosis; Phosphorylation; Protein tyrosine kinase A PtkA