cDNA cloning and expression of the flavoprotein d -aspartate oxidase from bovine kidney cortex
| Autor: | Stefano Duga, Tatjana Simonic, Massimo Malcovati, Armando Negri, Gabriella Tedeschi, Severino Ronchi, Maria Luisa Tenchini |
|---|---|
| Rok vydání: | 1997 |
| Předmět: |
D-aspartate oxidase
D-Aspartate Oxidase DNA Complementary Kidney Cortex Polyadenylation Molecular Sequence Data Flavoprotein Biochemistry Complementary DNA Animals Amino Acid Sequence Cloning Molecular Molecular Biology chemistry.chemical_classification Oxidase test Base Sequence biology Peroxisomal Targeting Signal 1 Cell Biology Molecular biology Amino acid Enzyme chemistry biology.protein Cattle Amino Acid Oxidoreductases Sequence Alignment Research Article |
| Zdroj: | Scopus-Elsevier Europe PubMed Central |
| ISSN: | 1470-8728 0264-6021 |
| DOI: | 10.1042/bj3220729 |
| Popis: | The isolation and sequencing of the complete cDNA coding for a d-aspartate oxidase, as well as the overexpression of the recombinant active enzyme, are reported for the first time. This 2022 bp cDNA, beside the coding portion, comprises a 5´ untranslated tract and the whole 3´ region including the polyadenylation signal and the poly(A) tail. The encoded protein comprises 341 amino acids, with the last three residues (-Ser-Lys-Leu) representing a peroxisomal targeting signal 1 (PTS1), hitherto unknown for this protein. The overexpression of recombinant d-aspartate oxidase was achieved in a prokaryotic system, and a soluble and active enzyme was obtained which accounted for about 10% of total bacterial protein. Comparisons with the known cDNAs for mammalian d-amino acid oxidase, another peroxisomal enzyme, are also made. The close structural and functional similarities shared by these enzymes at the protein level are not reflected at the nucleic acid level. |
| Databáze: | OpenAIRE |
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