Abstract:
:Mycobacterium tuberculosis acetohydroxyacid synthase (M. tuberculosis AHAS) has been proposed to bean essential target for novel herbicide- and chemical-based antibacterial agents. Therefore, here we investigated the roles of multiple conserved herbicide-binding site residues (R318, A146, Q148, M512, and V513) in M. tuberculosis AHAS through site-directed mutagenesis by characterizing the kinetic parameters and herbicide sensitivities of various point mutants. Interestingly, all mutant enzymes showed significantly altered kinetic parameters, specifically reduced affinity towards both the substrate and cofactor. Importantly, mutation of R318 led to a complete loss of AHAS activity, indicating a key role for this residue in substrate binding. Furthermore, all mutants demonstrated significant herbicide resistance against chlorimuron ethyl (CE), with several-fold higher IC50 than that of wild type AHAS. Docking analysis also indicated that binding of CE was slightly affected upon mutation of these residues. Taken together, these data suggest that the residues examined here mediate CE binding and may also be important for the catalytic activity of AHAS. This study will pave the way for future structure-function studies of CE and will also aid the development of novel anti-tuberculosis agents based on this chemical scaffold.
journal_name
Enzyme Microb Technoljournal_title
Enzyme and microbial technologyauthors
Jung IP,Cho JH,Koo BS,Yoon MYdoi
10.1016/j.enzmictec.2015.06.009subject
Has Abstractpub_date
2015-10-01 00:00:00pages
18-26eissn
0141-0229issn
1879-0909pii
S0141-0229(15)30014-4journal_volume
78pub_type
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journal_title:Enzyme and microbial technology
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journal_title:Enzyme and microbial technology
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