Structural and functional characterization of an arylamine N-acetyltransferase from the pathogen Mycobacterium abscessus: differences from other mycobacterial isoforms and implications for selective inhibition

• Published in: Acta crystallographica. Section D, Biological crystallography. Biological crystallography., 2014-11, Vol.70 (11), p.3066-3079
• Main Authors: Cocaign, Angélique, Kubiak, Xavier, Xu, Ximing, Garnier, Guillaume, Li de la Sierra-Gallay, Inès, Chi-Bui, Linh, Dairou, Julien, Busi, Florent, Abuhammad, Areej, Haouz, Ahmed, Dupret, Jean-Marie, Herrmann, Jean-Louis, Rodrigues-Lima, Fernando
• Format: Article
• Language: English
• Subjects: Acetylation; Amines; Amino Acid Sequence; Antibiotics; arylamine N-acetyltransferase; Arylamine N-Acetyltransferase - chemistry; Arylamine N-Acetyltransferase - genetics; Arylamine N-Acetyltransferase - metabolism; Crystallography, X-Ray; Enzymes; Humans; Inhibitors; Models, Molecular; Molecular Sequence Data; Mycobacterium; Mycobacterium - chemistry; Mycobacterium - enzymology; Mycobacterium - genetics; Mycobacterium - metabolism; Mycobacterium abscessus; Mycobacterium Infections - microbiology; Nocardia farcinica; Organic compounds; Pathogens; Phylogeny; Structural analysis; Substrate inhibition; Substrate Specificity
• ISSN: 1399-0047; 0907-4449; 1399-0047
• DOI: 10.1107/S1399004714021282

Mycobacterium abscessus is the most pathogenic rapid‐growing mycobacterium and is one of the most resistant organisms to chemotherapeutic agents. However, structural and functional studies of M. abscessus proteins that could modify/inactivate antibiotics remain nonexistent. Here, the structural and functional characterization of an arylamine N‐acetyltransferase (NAT) from M. abscessus [(MYCAB)NAT1] are reported. This novel prokaryotic NAT displays significant N‐acetyltransferase activity towards aromatic substrates, including antibiotics such as isoniazid and p‐aminosalicylate. The enzyme is endogenously expressed and functional in both the rough and smooth M. abscessus morphotypes. The crystal structure of (MYCAB)NAT1 at 1.8 Å resolution reveals that it is more closely related to Nocardia farcinica NAT than to mycobacterial isoforms. In particular, structural and physicochemical differences from other mycobacterial NATs were found in the active site. Peculiarities of (MYCAB)NAT1 were further supported by kinetic and docking studies showing that the enzyme was poorly inhibited by the piperidinol inhibitor of mycobacterial NATs. This study describes the first structure of an antibiotic‐modifying enzyme from M. abscessus and provides bases to better understand the substrate/inhibitor‐binding specificities among mycobacterial NATs and to identify/optimize specific inhibitors. These data should also contribute to the understanding of the mechanisms that are responsible for the pathogenicity and extensive chemotherapeutic resistance of M. abscessus.