title: High l-Trp affinity of indoleamine 2,3-dioxygenase 1 is attributed to two residues located in the distal heme pocket.
abstract: Indoleamine 2, 3-dioxygenase (IDO) catalyzes the oxidative cleavage of the pyrrole ring of l-Trp to generate N-formyl-kynurenine. Two IDO genes, IDO1 and IDO2, are found in vertebrates. Mammalian IDO1s are high-affinity, l-Trp-degrading enzymes, whereas IDO2s generally have a relatively low affinity. It has been suggested that the distal-Ser (corresponding to Ser167 of human IDO1) was crucial for improvement in the affinity for l-Trp but this idea was insufficient to explain the high affinity shown by mammalian IDO1. In this study, the amino acid sequences of vertebrate ancestral IDO1 and ancestral IDO2 were inferred, and bacterially expressed ancestral IDOs were characterized. Although the amino acid sequences of the enzymes shared high identity (86%) with each other, they showed distinct enzymatic properties. In analyses of a series of ancestral IDO1/IDO2 chimeric enzymes and their variants, the distal-Tyr (corresponding to Tyr126 of human IDO1) was detected as another and was probably the most crucial residue for high l-Trp affinity. The two amino acid substitutions (distal-Ser to Thr and distal-Tyr to His) drastically decreased the l-Trp affinity and catalytic efficiency of IDO1s. Conversely, two substitutions (distal-Thr to Ser and distal-His to Tyr) were sufficient to bestow IDO1-like high affinity on ancestral and chicken IDO2.
authors: Laboratory of Biochemistry, Department of Applied Science, Faculty of Science, National University Corporation Kochi University, Japan. julie@kochi-u.ac.jp.
authors: Hajime J
authors: HJ
authors: Yuasa
mesh_terms: D000596:Amino Acids / Q000737:chemistry; D000818:Animals; D020134:Catalytic Domain; D019143:Evolution, Molecular; D006418:Heme / Q000737:chemistry* / Q000378:metabolism; D006801:Humans; D050503:Indoleamine-Pyrrole 2,3,-Dioxygenase / Q000737:chemistry* / Q000145:classification / Q000378:metabolism; D016415:Sequence Alignment; D020539:Sequence Analysis, Protein; D014364:Tryptophan / Q000378:metabolism*; D014366:Tryptophan Oxygenase / Q000737:chemistry
chemical_list: D000596:Amino Acids; C576710:IDO1 protein, human; D050503:Indoleamine-Pyrrole 2,3,-Dioxygenase; D006418:Heme; D014364:Tryptophan; D014366:Tryptophan Oxygenase
keywords: ancestral enzyme; chimeric enzyme; indoleamine 2,3-dioxygenase; molecular evolution; tryptophan metabolism