생명과학회지 (Journal of Life Science)

  • 제18권8호
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  • Pages.1059-1065
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  • 2008
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  • 1225-9918(pISSN)
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  • 2287-3406(eISSN)
한국생명과학회 (Korean Society of Life Science)
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Kinesin superfamily KIF21A와 직접 결합하는 Pcp-2의 규명

Pcp-2 Interacts Directly with Kinesin Superfamily KIF21A Protein

  • 박혜영 (인제대학교 의과대학 생화학교실) ;
  • 김상진 (인제대학교 의과대학 신경과학교실) ;
  • 예성수 (인제대학교 의과대학 생화학교실) ;
  • 장원희 (인제대학교 의과대학 생화학교실) ;
  • 이상경 (인제대학교 의과대학 정신과학교실) ;
  • 박영홍 (인제대학교 의과대학 생화학교실) ;
  • 정용욱 (동국대학교 의과대학 해부학교실) ;
  • 문일수 (동국대학교 의과대학 해부학교실) ;
  • 김무성 (인제대학교 의과대학 신경외과학교실) ;
  • 석대현 (인제대학교 의과대학 생화학교실)
  • Park, Hye-Young (Departments of Biochemistry, College of Medicine, Inje University) ;
  • Kim, Sang-Jin (Departments of Neurology, College of Medicine, Inje University) ;
  • Ye, Sung-Su (Departments of Biochemistry, College of Medicine, Inje University) ;
  • Jang, Won-Hee (Departments of Biochemistry, College of Medicine, Inje University) ;
  • Lee, Sang-Kyeong (Departments of Psychiatry, College of Medicine, Inje University) ;
  • Park, Yeong-Hong (Departments of Biochemistry, College of Medicine, Inje University) ;
  • Jung, Yong-Wook (Departments of Anatomy, College of Medicine, Dongguk University) ;
  • Moon, Il-Soo (Departments of Anatomy, College of Medicine, Dongguk University) ;
  • Kim, Moo-Seong (Departments of Neurosurgery, College of Medicine, Inje University) ;
  • Seog, Dae-Hyun (Departments of Biochemistry, College of Medicine, Inje University)
  • 발행 : 2008.08.30
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초록

KIF21A는 kinesin superfamily에 속하는 분자 motor로서 미세소관을 따라서 분비소포를 운반한다. 최근의 연구결과 KIF21A 유전자 일부의 missense 돌연변이에 의하여 congenital fibrosis of the extraocular muscles (CFEOM) 1의 유발됨이 밝혀졌다. CFEOM1은 KIF21A의 돌연변이로 인하여 분화 발생과정에 occulo-motor신경과 neuromuscular junction 형성에 필요한 단백질을 이동시키지 못함으로써 유발된다. 본 연구에서는 효모 two-hybrid system을 사용하여 KIF21A의 WD-40 repeat domain과 결합하는 분자량이 작은 Purkinje cell protein-1 (Pcp-2), 또는 L7으로도 알려진 단백질을 분리하였다. Pcp-2는 효모 two-hybrid assay에서 KIF21A와 KIF21B의 WD-40 영역과는 결합하지만 다른 종류의 KIFs와는 결합하지 않았다. 또한 단백질간의 특이적 결합을 pull-down assay로 확인하였으며, 생쥐의 뇌 파쇄액에 Pcp-2 항체로 면역침강을 행하여 KIF21A를 확인한 결과 Pcp-2와 같이 침강하였다. 이러한 결과들은 KIF21A는 Pcp-2와 결합하며, 또한 Pcp-2는 KIF21A의 adaptor 단백질로서 세포 내 KIF21A의 수송에서 매개 단백질로 작용함을 시사한다.

KIF21A is a member of the Kinesin superfamily proteins (KIFs), which are microtubule-dependent molecular motors, anterograde axonal transporters of cargoes. Recently, congenital fibrosis of the extraocular muscles 1 (CFEOM1) has been shown to result from a small number of recurrent heterozygous missense mutations of KIF21A. CFEOM1 results from the inability of mutated KIF21A to successfully deliver cargoes to the development of the occulo-motor neuron or neuromuscular junction. Here, we used an yeast two-hybrid system to identify a protein that interacts with the WD-40 repeat domain of KIF21A and found a specific interaction with Purkinje cell protein-2 (Pcp-2), a small protein also known as L7. Pcp-2 protein bound to the WD-40 domain of KIF21A and KIF21B but not to other KIFs in yeast two-hybrid assays. In addition, this specific interaction was also observed in the glutathione S-transferase pull-down assay. An antibody to Pcp-2 specifically co-immunoprecipitated KIF21A associated with Pcp-2 from mouse brain extracts. These results suggest that Pcp-2 may be involved in the KIF21A-mediated transport as a KIF21A adaptor protein.

키워드

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