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Purification and sequence identification of anserinase
Shoji Yamada1, Yoshito Tanaka, Seiichi Ando
1Faculty of Fisheries, Kagoshima University, Japan.
The FEBS Journal
|November 24, 2005
Summary
Researchers identified the anserinase gene, a dipeptidase crucial for Nalpha-acetylhistidine hydrolysis in vertebrates. This discovery advances our understanding of metallopeptidases and their roles in various tissues.
Area of Science:
- Biochemistry
- Molecular Biology
- Enzymology
Background:
- Anserinase (Xaa-methyl-His dipeptidase, EC 3.4.13.5) is a key dipeptidase in poikilothermic vertebrates, hydrolyzing Nalpha-acetylhistidine.
- The specific gene encoding anserinase has remained uncharacterized until now.
Purpose of the Study:
- To molecularly identify and characterize the gene encoding anserinase.
- To investigate the tissue distribution of anserinase and related CNDP-like proteins.
Main Methods:
- Purification of anserinase from Nile tilapia (Oreochromis niloticus) brain.
- N-terminal sequencing of purified anserinase to design PCR primers.
- Cloning of anserinase cDNA and a related CNDP-like protein cDNA.
- Analysis of anserinase and CNDP-like protein mRNA tissue distribution via RT-PCR.
Main Results:
- The anserinase gene was molecularly identified, with its cDNA encoding a 476-amino acid mature protein (53.3 kDa) and an 18-amino acid signal peptide.
- Sequence analysis places anserinase within the M20A metallopeptidase subfamily, alongside serum carnosinase and cytosolic non-specific dipeptidase (CNDP).
- Anserinase mRNA was detected in brain, retina, kidney, and skeletal muscle, while CNDP-like protein mRNA was found in all examined tissues.
Conclusions:
- The molecular identification of anserinase provides a basis for further functional and structural studies.
- Anserinase and CNDP-like proteins exhibit distinct tissue expression patterns, suggesting specialized roles.
- This research contributes to the understanding of metallopeptidase diversity and function in vertebrates.