Structural analysis of mycobacterial branched-chain aminotransferase: implications for inhibitor design.
The branched-chain aminotransferase (BCAT) of Mycobacterium tuberculosis has been characterized as being essential to the survival of the bacterium. The enzyme is pyridoxal 5'-phosphate-dependent and belongs to the aminotransferase IIIa subfamily, to which the human BCATs also belong. The overall sequence similarity is high within the subfamily and the sequence identity among the active-site residues is high. In order to identify structurally unique features of M. tuberculosis BCAT, X-ray structural and functional analyses of the closely related BCAT from M. smegmatis were carried out. The crystal structures include the apo form at 2.2 A resolution and a 1.9 A structure of the holo form cocrystallized with the inhibitor O-benzylhydroxylamine (Obe). The analyses highlighted the active-site residues Tyr209 and Gly243 as being structurally unique characteristics of the mycobacterial BCATs relative to the human BCATs. The inhibitory activities of Obe and ammonium sulfate were verified in an inhibition assay. Modelling of the inhibitor Obe in the substrate pocket indicated potential for the design of a mycobacterial-specific inhibitor.
Castell
Alina
A
Department of Cell and Molecular Biology, Uppsala University, BMC, SE-751 24 Uppsala, Sweden.
Mille
Christian
C
Unge
Torsten
T
D000595:Amino Acid Sequence; D020134:Catalytic Domain; D018360:Crystallography, X-Ray; D004791:Enzyme Inhibitors / Q000737:chemistry* / Q000378:metabolism; D008958:Models, Molecular; D008969:Molecular Sequence Data; D009161:Mycobacterium / Q000201:enzymology*; D009169:Mycobacterium tuberculosis / Q000201:enzymology; D011487:Protein Conformation; D016415:Sequence Alignment; D000637:Transaminases / Q000037:antagonists & inhibitors* / Q000737:chemistry* / Q000378:metabolism
D004791:Enzyme Inhibitors; D000637:Transaminases; C019798:branched-chain-amino-acid transaminase