Purification and molecular cloning of a novel serine protease from the centipede, Scolopendra subspinipes mutilans

Weon Kyoo You, Young Doug Sohn, Ki Yong Kim, Doo Hong Park, Yangsoo Jang, Kwang Hoe Chung

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42 Citations (Scopus)

Abstract

A novel serine protease, named as scolonase, was purified and characterized from the tissue of the Korean centipede, Scolopendra subspinipes mutilans. Purified scolonase showed an apparent molecular weight of 25 kDa on sodium dodecyl sulfate-polyacrylamide gel electrophoresis analysis and an isoelectric point of 4.8 on isoelectric focusing gel. Scolonase was able to preferentially hydrolyze arginine over lysine at the cleavage site among the several synthetic peptide substrates. Scolonase has also a potent fibrinolytic activity by converting human Glu-plasminogen to activated plasmin due to the specific cleavage of the molecule at the peptide bond Arg561-Val 562. The enzyme activity of scolonase was completely inhibited by phenylmethanesulfonyl fluoride and difluorophosphate. The cDNA encoding scolonase was cloned from the cDNA library of the centipede constructed with oligonucleotide probe, which was designed on the basis of the N-terminal amino acid sequence of scolonase. The deduced complete amino acid sequence of scolonase demonstrated that the protein is composed of 277 amino acids including 33 amino acids as a leader sequence, and that it has significant sequence homology with other serine proteases.

Original languageEnglish
Pages (from-to)239-250
Number of pages12
JournalInsect Biochemistry and Molecular Biology
Volume34
Issue number3
DOIs
Publication statusPublished - 2004 Mar

Bibliographical note

Funding Information:
We gratefully acknowledge the help and advice of Dr. J.H. Yoon on the molecular structure of scolonase and of other serine proteases by computer modeling, and we thank Dr. H.M. Moon and S.I. Chung for their continued support. We also thank In2gen Co., Ltd. (Seoul, Korea) for the MS/MS spectroscopy analysis. This work was supported in part by the Korea Green Cross Corp and by G7 grant from the Korean Ministry of Science and Technology.

All Science Journal Classification (ASJC) codes

  • Biochemistry
  • Molecular Biology
  • Insect Science

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