3-hydroxy-kynurenine transaminase identity with alanine glyoxylate transaminase - A probable detoxification protein in Aedes aegypti
Autor(es): Han, Q. ; Fang, J. M. ; Li, J. Y.
Resumo: This study describes the functional characterization of a specific mosquito transaminase responsible for catalyzing the transamination. of 3-hydroxykynurenine (3-HK) to xanthurenic acid (XA). The enzyme was purified from Aedes aegypti larvae by ammonium sulfate fractionation, heat treatment, and various chromatographic techniques, plus non-denaturing electrophoresis. The purified transaminase has a relative molecular mass of 42,500 by SDS-PAGE. N-terminal and internal sequencing of the purified protein and its tryptic fragments resolved a partial N-terminal sequence of 19 amino acid residues and 3 partial internal peptide sequences with 7, 10, and 7 amino acid residues. Using degenerate primers based on the partial internal sequences for PCR amplification and cDNA library screening, a full-length cDNA clone with a 1,167-bp open reading frame was isolated. Its deduced amino acid sequence consists of 389 amino acid residues with a predicted molecular mass of 43,239 and shares 45-46% sequence identity with mammalian alanine glyoxylate transaminases. Northern analysis shows the active transcription of the enzyme in larvae and developing eggs. Substrate specificity analysis of this mosquito transaminase demonstrates that the enzyme is active with 3-HK, kynurenine, or alanine substrates. The enzyme has greater affinity and catalytic efficiency for 3-HK than for kynurenine and alanine. The biochemical characteristics of the enzyme in conjunction with the profiles of 3-HK transaminase, activity and XA accumulation during mosquito development clearly point out its physiological function in the 3-HK to XA pathway. Our data suggest that the mosquito transaminase was evolved in a manner precisely reflecting the physiological requirement of detoxifying 3-HK produced in the tryptophan oxidation pathway in the mosquito.
Palavras-Chave: Major branch pathway ; Neuronal cell-death ; Xanthurenic acid ; Kynurenine aminotransferase ; Endogenous neurotoxin ; Huntingtons-disease ; Frontal-cortex ; Rat-brain ; 3-Hydroxykynurenine ; Mosquito
Imprenta: The Journal of Biological Chemistry, v. 277, p. 15781-15787, 2002
Identificador do objeto digital: 10.1074/jbc.M201202200
Descritores: Aedes aegypti - Cell ; Aedes aegypti - Proteins ; Aedes aegypti - PCR detection ; Aedes aegypti - Molecular screening
Data de publicação: 2002
