생명과학회지 (Journal of Life Science)

  • 제21권8호
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  • Pages.1076-1082
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  • 2011
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  • 1225-9918(pISSN)
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  • 2287-3406(eISSN)
한국생명과학회 (Korean Society of Life Science)
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KIF5s와 직접 결합하는 액틴 결합 운동단백질 Myo9s의 규명

Direct Interaction of KIF5s and Actin-Based Transport Motor, Myo9s

  • 석대현 (인제대학교 의과대학 생화학교실)
  • Seog, Dae-Hyun (Department of Biochemistry, College of Medicine, Inje University)
  • 투고 : 2011.05.12
  • 심사 : 2011.07.12
  • 발행 : 2011.08.30
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초록

미세소관(microtubule) 위를 이동하는 키네신은 분비소포를 이동시키는 운동단백질이다. KIF5s (KIF5A, KIF5B and KIF5C)는 세포막으로 싸인 각종 세포 내 소기관과 결합하여 미세소관을 따라 목적지까지 이동시킨다는 결과는 알려져 있지만, 어떻게 상대의 cargo를 인식하는지는 밝혀지지 않았다. 본 연구는 KIF5B의 결합 단백질을 동정하기 위하여 효모 two-hybrid system을 사용하여 KIF5B와 특이적으로 결합하는 Myo9b을 확인하였다. Myo9b는 액틴위를 이동하는 운동단백질로 다른 KIF5s들과도 결합함을 효모 two-hybrid assay로 확인하였다. 또한 Myo9s의 GTPase 활성화 단백질(GAP) 영역은 KIF5B와 결합하는데 필수영역임을 확인하였고, 이러한 단백질간의 결합은 Glutathione S-transferase (GST) pull-down assay를 통하여서도 확인하였다. 생쥐의 뇌 파쇄액에 KIF5B들의 항체로 면역침강을 행하여 Myo9s 단백질을 확인한 결과, KIF5s는 Myo9s 단백질과 특이적으로 함께 침강하였다. 이러한 결과들은 kinesin-I는 액틴 결합 운동단백질과 직접 결합함을 보여준다.

Microtubule-based kinesin motor proteins are used for long-range vesicular transport. KIF5s (KIF5A, KIF5B and KIF5C) mediate the transport of various membranous vesicles along microtubules, but the mechanism behind how they recognize and bind to a specific cargo has not yet been completely elucidated. To identify the interaction protein for KIF5B, yeast two-hybrid screening was performed and a specific interaction with the unconventional myosin Myo9b, an actin-based vesicle transport motor, was found. The GTPase-activating protein (GAP) domain of Myo9s was essential for interaction with KIF5B in the yeast two-hybrid assay. Myo9b bound to the carboxyl-terminal region of KIF5B and to other KIF5 members. In addition, glutathione S-transferase (GST) pull-downs showed that Myo9s specifically interact to the complete Kinesin-I complex. An antibody to KIF5B specifically co-immunoprecipitated KIF5B associated with Myo9s from mouse brain extracts. These results suggest that kinesin-I motor protein interacts directly with actin-based motor proteins in the cell.

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