cDNA cloning and expression of the flavoprotein D-aspartate oxidase from bovine kidney cortex

Tatjana Simonic, Stefano Duga, Armando Negri, Gabriella Tedeschi, Massimo Malcovati, Maria L. Tenchini, Severino Ronchi

Research output: Contribution to journalArticle

36 Citations (Scopus)

Abstract

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.

Original languageEnglish
Pages (from-to)729-735
Number of pages7
JournalBiochemical Journal
Volume322
Issue number3
Publication statusPublished - Mar 15 1997

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D-Aspartate Oxidase
Flavoproteins
Kidney Cortex
Cloning
Organism Cloning
Complementary DNA
Enzymes
seryl-lysyl-leucine
D-Amino-Acid Oxidase
Polyadenylation
Proteins
Bacterial Proteins
Nucleic Acids
Amino Acids
Messenger RNA

ASJC Scopus subject areas

  • Biochemistry

Cite this

Simonic, T., Duga, S., Negri, A., Tedeschi, G., Malcovati, M., Tenchini, M. L., & Ronchi, S. (1997). cDNA cloning and expression of the flavoprotein D-aspartate oxidase from bovine kidney cortex. Biochemical Journal, 322(3), 729-735.

cDNA cloning and expression of the flavoprotein D-aspartate oxidase from bovine kidney cortex. / Simonic, Tatjana; Duga, Stefano; Negri, Armando; Tedeschi, Gabriella; Malcovati, Massimo; Tenchini, Maria L.; Ronchi, Severino.

In: Biochemical Journal, Vol. 322, No. 3, 15.03.1997, p. 729-735.

Research output: Contribution to journalArticle

Simonic, T, Duga, S, Negri, A, Tedeschi, G, Malcovati, M, Tenchini, ML & Ronchi, S 1997, 'cDNA cloning and expression of the flavoprotein D-aspartate oxidase from bovine kidney cortex', Biochemical Journal, vol. 322, no. 3, pp. 729-735.
Simonic T, Duga S, Negri A, Tedeschi G, Malcovati M, Tenchini ML et al. cDNA cloning and expression of the flavoprotein D-aspartate oxidase from bovine kidney cortex. Biochemical Journal. 1997 Mar 15;322(3):729-735.
Simonic, Tatjana ; Duga, Stefano ; Negri, Armando ; Tedeschi, Gabriella ; Malcovati, Massimo ; Tenchini, Maria L. ; Ronchi, Severino. / cDNA cloning and expression of the flavoprotein D-aspartate oxidase from bovine kidney cortex. In: Biochemical Journal. 1997 ; Vol. 322, No. 3. pp. 729-735.
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