title: Human mitochondrial branched chain aminotransferase: structural basis for substrate specificity and role of redox active cysteines.
abstract: Crystal structures of the fold type IV pyridoxal phosphate (PLP)-dependent human mitochondrial branched chain aminotransferase (hBCATm) reaction intermediates have provided a structural explanation for the kinetically determined substrate specificity of hBCATm. The isoleucine side chain in the ketimine intermediate occupies a hydrophobic binding pocket that can be defined by three surfaces. Modeling of amino acids on the ketimine structure shows that the side chains of nonsubstrate amino acids such as the aromatic amino acids, alanine, or aspartate either are unable to interact through van der Waals' interactions or have steric clashes. The structural and biochemical basis for the sensitivity of the mammalian BCAT to reducing agents has also been elucidated. Two cysteine residues in hBCATm, Cys315 and Cys318 (CXXC), are part of a redox-controlled mechanism that can regulate hBCATm activity. The residues surrounding Cys315 and Cys318 show considerable sequence conservation in the prokaryotic and eukaryotic BCAT sequences, however, the CXXC motif is found only in the mammalian proteins. The results suggest that the BCAT enzymes may join the list of enzymes that can be regulated by redox status.
authors.affiliation: Department of Biochemistry, Wake Forest University School of Medicine, Medical Center Boulevard, Winston-Salem, NC 27157, USA.
authors.forename: Myra E
authors.initials: ME
authors.lastname: Conway
authors.forename: Neela
authors.initials: N
authors.lastname: Yennawar
authors.forename: Reidar
authors.initials: R
authors.lastname: Wallin
authors.forename: Leslie B
authors.initials: LB
authors.lastname: Poole
authors.forename: Susan M
authors.initials: SM
authors.lastname: Hutson
mesh_terms: D000595:Amino Acid Sequence; D003545:Cysteine / Q000378:metabolism*; D006801:Humans; D008928:Mitochondria / Q000201:enzymology*; D008969:Molecular Sequence Data; D010084:Oxidation-Reduction; D013379:Substrate Specificity; D000637:Transaminases / Q000737:chemistry / Q000378:metabolism*
chemical_list: D000637:Transaminases; C019798:branched-chain-amino-acid transaminase; D003545:Cysteine