Biochemical characterization of zebrafish Prss59.1

Rubel Rana; Forhad Hossain; Hasan Ali; Maisum Sarwar Jyoti; Toshinobu Tokumoto

doi:10.1016/j.bbrc.2021.04.118

Biochemical characterization of zebrafish Prss59.1

Biochem Biophys Res Commun. 2021 Jun 30:560:32-36. doi: 10.1016/j.bbrc.2021.04.118. Epub 2021 May 6.

Authors

Rubel Rana¹, Forhad Hossain², Hasan Ali¹, Maisum Sarwar Jyoti¹, Toshinobu Tokumoto³

Affiliations

¹ Integrated Bioscience Section, Graduate School of Science and Technology, National University Corporation, Shizuoka University, 836 Ohya, Suruga-ku, Shizuoka, 422-8529, Japan.
² Biological Science Course, Graduate School of Science, National University Corporation, Shizuoka University, Oya 836, Suruga-ku, Shizuoka, 422-8529, Japan.
³ Integrated Bioscience Section, Graduate School of Science and Technology, National University Corporation, Shizuoka University, 836 Ohya, Suruga-ku, Shizuoka, 422-8529, Japan; Biological Science Course, Graduate School of Science, National University Corporation, Shizuoka University, Oya 836, Suruga-ku, Shizuoka, 422-8529, Japan. Electronic address: tokumoto.toshinobu@shizuoka.ac.jp.

PMID: 33965786
DOI: 10.1016/j.bbrc.2021.04.118

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.

Keywords: Peptidase; Recombinant protein; Renature; Trypsin; Zebrafish; prss59.1.

Publication types

Research Support, Non-U.S. Gov't

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*

Substances

Zebrafish Proteins
Peptide Hydrolases