Inhibition of 7,8-diaminopelargonic acid aminotransferase by amiclenomycin and analogues.
Cis and trans stereoisomers of amiclenomycin, a natural L-amino acid antibiotic, have been prepared using unequivocal routes. By using 1H NMR spectroscopy, the configuration of the six-membered ring of natural amiclenomycin was shown to be cis and not trans as originally proposed. Amiclenomycin and some synthetic analogues with the cis configuration irreversibly inactivate DAPA AT (7,8-diaminopelargonic acid aminotransferase), an enzyme involved in biotin biosynthesis, by forming an aromatic PLP (pyridoxal-5'-phosphate)-inhibitor adduct that is tightly bound to the active site. The following kinetic parameters for the inactivation of Escherichia coli DAPA AT by amiclenomycin were derived: K(I)=2 microM and k(inact)=0.4 min(-1). The structure of the aromatic adduct formed upon inactivation was confirmed by UV-visible spectroscopy, X-ray crystal structure determination and MS. Because Mycobacterium tuberculosis DAPA AT is a potential drug target, this enzyme was cloned, overexpressed and purified to homogeneity for biochemical characterization.
Mann
S
S
Synthèse, Structure et Fonction de Molécules Bioactives, UMR CNRS 7613, Université Pierre et Marie Curie, boîte 182, 4, place Jussieu, 75252 Paris Cedex 05, France. mann@ccr.jussieu.fr
Marquet
A
A
Ploux
O
O
D000613:Aminobutyrates / Q000494:pharmacology*; D000995:Antitubercular Agents / Q000494:pharmacology; D001426:Bacterial Proteins / Q000037:antagonists & inhibitors; D004791:Enzyme Inhibitors / Q000494:pharmacology*; D007700:Kinetics; D009169:Mycobacterium tuberculosis / Q000201:enzymology; D000637:Transaminases / Q000037:antagonists & inhibitors*
D000613:Aminobutyrates; D000995:Antitubercular Agents; D001426:Bacterial Proteins; D004791:Enzyme Inhibitors; C006488:amiclenomycin; C020029:7,8-diaminopelargonic acid aminotransferase; D000637:Transaminases