Molecular basis for the inhibition of β-hydroxyacyl-ACP dehydratase HadAB complex from Mycobacterium tuberculosis by flavonoid inhibitors.
10.1007/s13238-015-0181-1
- Author:
Yu DONG
1
;
Xiaodi QIU
;
Neil SHAW
;
Yueyang XU
;
Yuna SUN
;
Xuemei LI
;
Jun LI
;
Zihe RAO
Author Information
1. National Laboratory of Biomacromolecules, Institute of Biophysics, Chinese Academy of Sciences, Beijing, 100101, China.
- Publication Type:Journal Article
- MeSH:
Amino Acid Sequence;
Bacterial Proteins;
chemistry;
metabolism;
Catalytic Domain;
Enzyme Inhibitors;
chemistry;
pharmacology;
Flavonoids;
chemistry;
pharmacology;
Hydro-Lyases;
antagonists & inhibitors;
chemistry;
Molecular Sequence Data;
Mycobacterium tuberculosis;
drug effects;
enzymology;
Protein Binding;
drug effects;
Protein Multimerization;
drug effects;
Protein Structure, Secondary;
Sequence Alignment
- From:
Protein & Cell
2015;6(7):504-517
- CountryChina
- Language:English
-
Abstract:
Dehydration is one of the key steps in the biosynthesis of mycolic acids and is vital to the growth of Mycobacterium tuberculosis (Mtb). Consequently, stalling dehydration cures tuberculosis (TB). Clinically used anti-TB drugs like thiacetazone (TAC) and isoxyl (ISO) as well as flavonoids inhibit the enzyme activity of the β-hydroxyacyl-ACP dehydratase HadAB complex. How this inhibition is exerted, has remained an enigma for years. Here, we describe the first crystal structures of the MtbHadAB complex bound with flavonoid inhibitor butein, 2',4,4'-trihydroxychalcone or fisetin. Despite sharing no sequence identity from Blast, HadA and HadB adopt a very similar hotdog fold. HadA forms a tight dimer with HadB in which the proteins are sitting side-by-side, but are oriented anti-parallel. While HadB contributes the catalytically critical His-Asp dyad, HadA binds the fatty acid substrate in a long channel. The atypical double hotdog fold with a single active site formed by MtbHadAB gives rise to a long, narrow cavity that vertically traverses the fatty acid binding channel. At the base of this cavity lies Cys61, which upon mutation to Ser confers drug-resistance in TB patients. We show that inhibitors bind in this cavity and protrude into the substrate binding channel. Thus, inhibitors of MtbHadAB exert their effect by occluding substrate from the active site. The unveiling of this mechanism of inhibition paves the way for accelerating development of next generation of anti-TB drugs.
