PUBLICATION
Identification of a puromycin-sensitive aminopeptidase from zebrafish (Danio rerio)
- Authors
- Chen, S.H., Cao, M.J., Huang, J.Z., and Wu, G.P.
- ID
- ZDB-PUB-110207-6
- Date
- 2011
- Source
- Comparative biochemistry and physiology. Part B, Biochemistry & molecular biology 159(1): 10-7 (Journal)
- Registered Authors
- Keywords
- Zebrafish, Puromycin-sensitive aminopeptidase, Purification, Characterization
- MeSH Terms
-
- Amino Acid Sequence
- Aminopeptidases/chemistry*
- Aminopeptidases/isolation & purification
- Aminopeptidases/metabolism
- Animals
- Molecular Sequence Data
- Protease Inhibitors/pharmacology
- Puromycin/pharmacology
- Substrate Specificity
- Zebrafish Proteins/chemistry*
- Zebrafish Proteins/isolation & purification
- Zebrafish Proteins/metabolism
- PubMed
- 21276865 Full text @ Comp. Biochem. Physiol. B Biochem. Mol. Biol.
Citation
Chen, S.H., Cao, M.J., Huang, J.Z., and Wu, G.P. (2011) Identification of a puromycin-sensitive aminopeptidase from zebrafish (Danio rerio). Comparative biochemistry and physiology. Part B, Biochemistry & molecular biology. 159(1):10-7.
Abstract
An aminopeptidase from zebrafish (Danio rerio) was purified 1,247-fold to homogeneity with 35.4% recovery by column chromatography successively on DEAE-Sephacel, hydroxyapatite, and Phenyl-Sepharose. The molecular mass of the enzyme was estimated as 98kDa by SDS-PAGE and gel filtration. Optimum temperature and pH of the enzyme were 45°C and 7.5, respectively. The enzyme preferentially hydrolyzed substrate Leu-MCA with k(cat)/K(m) of 4.2×10(6) M(-1)s(-1) and activation energy of 80.5kJ·M(-1), respectively. It was specifically inhibited by bestatin, puromycin and metal-chelating agents, and Zn(2+) seemed to be its metal cofactor(s). Some L-amino acids significantly inhibited its activity, and L-cysteine was a non-competitive inhibitor with a K(i) of 0.27mM. According to the peptide mass fingerprint analysis, the enzyme was highly matched with the predicted Danio rerio aminopeptidase puromycin sensitive (gi:255683530) (EC 3.4.11.14), suggesting the present enzyme is a puromycin-sensitive aminopeptidase of zebrafish.
Genes / Markers
Expression
Phenotype
Mutations / Transgenics
Human Disease / Model
Sequence Targeting Reagents
Fish
Orthology
Engineered Foreign Genes
Mapping