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Roles of Src-family kinase isoforms, Lyn, Fyn, Fgr, and c-Src on degranulation in RBL-2H3 mast cells

비만세포의 탈과립에 대한 다양한 Src-family kinase의 역할

  • Lee, Jun-Ho (Department of Immunology, College of Medicine, Konkuk University) ;
  • Mun, Se-Hwan (Department of Immunology, College of Medicine, Konkuk University) ;
  • Ko, Na-Young (Department of Immunology, College of Medicine, Konkuk University) ;
  • Kim, Jie-Wan (Department of Immunology, College of Medicine, Konkuk University) ;
  • Kim, Do-Kyun (Department of Immunology, College of Medicine, Konkuk University) ;
  • Kim, Joo-Dong (Department of Immunology, College of Medicine, Konkuk University) ;
  • Her, Erk (Department of Immunology, College of Medicine, Konkuk University) ;
  • Choi, Wahn-Soo (Department of Immunology, College of Medicine, Konkuk University)
  • 이준호 (건국대학교 의학전문대학원 면역학교실) ;
  • 문세환 (건국대학교 의학전문대학원 면역학교실) ;
  • 고나영 (건국대학교 의학전문대학원 면역학교실) ;
  • 김지완 (건국대학교 의학전문대학원 면역학교실) ;
  • 김도균 (건국대학교 의학전문대학원 면역학교실) ;
  • 김주동 (건국대학교 의학전문대학원 면역학교실) ;
  • 허억 (건국대학교 의학전문대학원 면역학교실) ;
  • 최완수 (건국대학교 의학전문대학원 면역학교실)
  • Published : 2007.03.30

Abstract

The rat RBL-2H3 mast cells contain various Src-family kinases. Previous reports with this cell line indicated that Lyn activation is an important initial signaling for the activation of the cells. However, the role and location of other Src-family kinase isoforms which are expressed in the cells are not clear. In this study, we now show that isoforms of Src-family kinases, Lyn, fyn, Fgr, c-Src, and Yes are differentially expressed and located differently in the cells as indicated by RT-PCR, immunoblotting analysis, and confocal microscopy. Lyn and Fgr were located on plasma membrane but on the other hand c-Src and Yes were located on intracellular organelle. All of Src-family kinases were cloned and overexpressed for investigating the roles of the isoforms. Overexpression of Fyn and Fgr, not Lyn and c-Src, stimulated Ag-induced degranulation in the cells. Our findings strongly suggest for the first time that each of Src-family kinase isoform can regulate differentially $Fc{\varepsilon}RI$-mediated signaling in RBL-2H3 mast cells.

흰쥐유래의 비만세포인 RBL-2H3 세포는 다양한 Src-family kinase를 발현한다. 현재까지의 연구결과에 의하면 비만세포의 초기 활성화에 Lyn kinase가 중요한 역할을 한다고 알려져 왔다. 그러나 그 세포에서 발현되는 다양한 다른 Src-family kinase의 역할은 불분명하다. 본 연구에서는 비만세포에서 다양한 Src-family kinase가 세포내 다른 곳에서 다양하게 발현되고 있다는 사실을 RT-PCR, immunoblotting 그리고 confocal microscopy 기법을 이용하여 증명하였다. 그 결과 Lyn 및 Fgr kinase는 세포막에 위치하고 c-Src 및 Yes kinase는 세포 내 과립에 존재하는 것을 알 수 있었다. 모든 Src-family kinase를 클로닝하고 과발현하여 탈과립 대한 영향을 평가하였다. 그 결과 fyn과 Fgr kinase는 비만세포에서 항원 유도의 탈과립을 증가시켰으며 반면 Lyn kinsae는 탈과립을 억제시키는 것을 확인할 수 있었다. 이러한 결과는 비만세포 초기 신호전달계에서 Fgr가 중요한 역할을 할 가능성을 제시한다.

Keywords

References

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