Cloning of a matrix metalloproteinase cDNA from Scylliorhinus torazame

두툽상어 matrix metalloproteinase 유전자 cDNA의 클로닝

  • 김종원 (부산대학교 자연과학대학 분자생물학과) ;
  • 조원진 (부산대학교 자연과학대학 분자생물학과) ;
  • 천광호 (부산대학교 자연과학대학 분자생물학과) ;
  • 김규원 (부산대학교 자연과학대학 분자생물학과) ;
  • 김영진 (부산대학교 자연과학대학 분자생물학과) ;
  • 이상준 (국립수산진흥원 생물공학과) ;
  • 신혜자 (동서대학교 환경공학과) ;
  • 임운기 (부산대학교 자연과학대학 분자생물학과)
  • Published : 1998.06.01

Abstract

Matrix metalloproteinases(MMPs) are a group of zinc enzymes responsible for degradation of the matrix components such as collagen and proteoglycans in normal embryogenesis and remodeling and in many disease processes such as arthritis, cancer, periodontitis, and osteprocess. Genetically distince MMPs have been characterized and their genes have been cloned thus far from a variaty of species but not from fishes. In this stydy, a mmp cDNA was cloned by using RT-PCR(reverse transcriptase dependent polymerase chain reaction) from Scylliorhinus toraxzame(shark), agroup of cartilaginous fish, abundant in the coast of Pusan, Korea. It has 74% base homologue with membrane type matrix matalloproteinase-3 genes(mt3-mmps) from human, rat and chick, and also shows more than 90% residue homologue with them. In addition, it has cysteine switch domain, zinc binding domain(HExGH motif), propeptide cleavage site, and RRKR motif, which are present in MMPs. This result indicates that cDNA fragment cloned here may be mt3-mmp or its analogous gejne cDNA fragment of Scylliorhinus torzame.

Matrix metalloproteinases(MMP)는 배발생 및 재조직화 등의 정상적인 생체형성과 관절염, 암전이, 치근막염, 골조송증 등의 질병과정에서 collagen이나 proteoglycan과 같은 세포외기질의 구성성분을 분해하는 아연(zinc)효소군이다. 지금까지 다양한 종에서 mmp의 유전자가 클로닝되고 그 기능이 연구되어 왔지만 아직 어류에서는 연구결과가 보고된 바가 없다. 본 연구에서는 한국의 부산연안에 많은 연골어유 투툽상어(Scylliorhinus toraxzame)로부터 RT-PCR(reverse transcriptase dependent polymerase chain reaction)의방법으로 mmp cDNA의 일부를 클로닝하였다. 이것은 염기서열에서 인간, 쥐 및 닭의 membrane type matrix matalloproteinase-3(mt3-mmps)의 염기서열과 74% 동일성을 보이며, 아미노산서열에서는 90%이상의 동일성을 갖고 있다. 또한 MMP에 나타나는 cysteine switch domain, zinc binding domain(HExGH motif), propeptide cleavage site, and RRKR motif등을 가지고 있다. 이러한 결과로부터 본 연구에서 클로닝된 RT-PCR단편은 두툽상어의 mt3-mmp 또는 이와 유사한 유전자의 cDNA이라 믿어진다.

Keywords

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