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Published on: May 9, 2015
Caenorhabditis elegans has two genes encoding functional d-aspartate oxidases.
Masumi Katane1, Yousuke Seida, Masae Sekine
1Laboratory of Biomolecular Science, School of Pharmaceutical Sciences, Kitasato University, Tokyo, Japan.
The FEBS Journal
|December 5, 2006
Summary
This study reports the first cDNA cloning of invertebrate d-amino acid oxidase (DAAO) and d-aspartate oxidase (DASPO) genes in Caenorhabditis elegans. Researchers identified one DAAO and two distinct DASPO genes, revealing novel insights into enzyme diversity.
Area of Science:
- Biochemistry
- Molecular Biology
- Genetics
Background:
- d-amino acid oxidase (DAAO) and d-aspartate oxidase (DASPO) enzymes play crucial roles in various biological processes.
- Previously, it was believed that organisms possess only one gene copy for each of these enzymes.
- Understanding the diversity and function of these enzymes in invertebrates is essential.
Purpose of the Study:
- To isolate and characterize cDNA clones encoding DAAO and DASPO from Caenorhabditis elegans.
- To investigate the enzymatic activities and substrate specificities of the expressed recombinant proteins.
- To determine the number of functional DAAO and DASPO genes in C. elegans.
Main Methods:
- RNA isolation and RT-PCR from Caenorhabditis elegans.
- cDNA cloning and expression of recombinant proteins in Escherichia coli with N-terminal His-tag.
- Biochemical assays to determine enzyme activity and substrate specificity of purified recombinant proteins.
Main Results:
- Four cDNA clones were isolated, encoding proteins Y69Ap, C47Ap, F18Ep, and F20Hp.
- Recombinant Y69Ap exhibited DAAO activity, deaminating neutral and basic d-amino acids.
- Recombinant C47Ap and F18Ep demonstrated DASPO activity, deaminating acidic d-amino acids, with distinct kinetic properties.
- F20Hp was not recovered in soluble fractions.
Conclusions:
- This study presents the first cDNA cloning of invertebrate DAAO and DASPO genes.
- Caenorhabditis elegans possesses at least two functional DASPO genes and one DAAO gene, challenging previous assumptions of single gene copies.
- The identified DASPO enzymes exhibit differing substrate specificities and potentially distinct subcellular localizations, highlighting enzyme diversity within a single organism.
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