title: Crystal structure of Saccharomyces cerevisiae Aro8, a putative α-aminoadipate aminotransferase.
abstract: α-Aminoadipate aminotransferase (AAA-AT) catalyzes the amination of 2-oxoadipate to α-aminoadipate in the fourth step of the α-aminoadipate pathway of lysine biosynthesis in fungi. The aromatic aminotransferase Aro8 has recently been identified as an AAA-AT in Saccharomyces cerevisiae. This enzyme displays broad substrate selectivity, utilizing several amino acids and 2-oxo acids as substrates. Here we report the 1.91Å resolution crystal structure of Aro8 and compare it to AAA-AT LysN from Thermus thermophilus and human kynurenine aminotransferase II. Inspection of the active site of Aro8 reveals asymmetric cofactor binding with lysine-pyridoxal-5-phosphate bound within the active site of one subunit in the Aro8 homodimer and pyridoxamine phosphate and a HEPES molecule bound to the other subunit. The HEPES buffer molecule binds within the substrate-binding site of Aro8, yielding insights into the mechanism by which it recognizes multiple substrates and how this recognition differs from other AAA-AT/kynurenine aminotransferases.
authors.affiliation: Department of Biological Chemistry, University of Michigan, Ann Arbor, Michigan, 48109.
authors.forename: Stacie L
authors.initials: SL
authors.lastname: Bulfer
authors.forename: Joseph S
authors.initials: JS
authors.lastname: Brunzelle
authors.forename: Raymond C
authors.initials: RC
authors.lastname: Trievel
mesh_terms: D051307:2-Aminoadipate Transaminase / Q000737:chemistry* / Q000378:metabolism; D020134:Catalytic Domain / Q000235:genetics; D018360:Crystallography, X-Ray; D006531:HEPES / Q000378:metabolism; D006801:Humans; D008239:Lysine / Q000378:metabolism; D008958:Models, Molecular; D017434:Protein Structure, Tertiary; D011732:Pyridoxal Phosphate / Q000378:metabolism; D011733:Pyridoxamine / Q000031:analogs & derivatives / Q000378:metabolism; D012441:Saccharomyces cerevisiae / Q000201:enzymology*; D029701:Saccharomyces cerevisiae Proteins / Q000737:chemistry* / Q000378:metabolism; D013379:Substrate Specificity; D016963:Thermus thermophilus / Q000201:enzymology*; D000637:Transaminases / Q000737:chemistry* / Q000378:metabolism
chemical_list: D029701:Saccharomyces cerevisiae Proteins; D011732:Pyridoxal Phosphate; D011733:Pyridoxamine; D000637:Transaminases; D051307:2-Aminoadipate Transaminase; C019729:aromatic amino acid aminotransferase; C021481:kynurenine-oxoglutarate transaminase; D008239:Lysine; C010627:pyridoxamine phosphate; D006531:HEPES
keywords: X-ray crystallography; aminotransferase; aromatic aminotransferase I; lysine biosynthesis; multi-substrate enzyme; pyridoxal 5′-phosphate
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