PUBLICATION
Biochemical characterization of zebrafish Prss59.1
- Authors
- Rana, R., Hossain, F., Ali, H., Sarwar Jyoti, M., Tokumoto, T.
- ID
- ZDB-PUB-210511-5
- Date
- 2021
- Source
- Biochemical and Biophysical Research Communications 560: 32-36 (Journal)
- Registered Authors
- Tokumoto, Toshinobu
- Keywords
- Peptidase, Recombinant protein, Renature, Trypsin, Zebrafish, prss59.1
- MeSH Terms
-
- Hydrogen-Ion Concentration
- Peptide Hydrolases/chemistry
- Peptide Hydrolases/genetics
- Peptide Hydrolases/isolation & purification
- Peptide Hydrolases/metabolism*
- Sequence Analysis, Protein
- Substrate Specificity
- Temperature
- Zebrafish Proteins/chemistry
- Zebrafish Proteins/genetics
- Zebrafish Proteins/isolation & purification
- Zebrafish Proteins/metabolism*
- PubMed
- 33965786 Full text @ Biochem. Biophys. Res. Commun.
Citation
Rana, R., Hossain, F., Ali, H., Sarwar Jyoti, M., Tokumoto, T. (2021) Biochemical characterization of zebrafish Prss59.1. Biochemical and Biophysical Research Communications. 560:32-36.
Abstract
Eleven genes, including prss59.1, were selected as candidate ovulation-inducing genes on the basis of microarray analysis and RNA sequencing in our previous study. To address the role of prss59.1, the prss59.1 gene knock-out zebrafish strain is currently being established by genome editing. In this study, for further phenotypic analysis of prss59.1, biochemical characterization of Prss59.1 was conducted using recombinant protein. A C-terminal histidine-tagged version of zebrafish Prss 59.1 was constructed. Although E. coli-produced recombinant Prss59.1 showed almost no activity, peptidase activities appeared after denaturation and renaturation. Zebrafish Prss59.1 showed the highest activity against Lys-MCA. The optimal temperature and pH of the activity toward Lys-MCA were 37 °C and pH 8.0, respectively. The Km value was 0.17 mM. Thus, zebrafish Prss59.1 possesses the closed character of trypsin, as expected from the DNA sequence.
Genes / Markers
Expression
Phenotype
Mutations / Transgenics
Human Disease / Model
Sequence Targeting Reagents
Fish
Orthology
Engineered Foreign Genes
Mapping