The role of kinesin neck region in force generation

• Project/Area Number: 08458204
• Research Category: Grant-in-Aid for Scientific Research (B)
• Allocation Type: Single-year Grants
• Section: 一般
• Research Field: Biophysics
• Research Institution: The University of Tokyo
• Principal Investigator: TOYOSHIMA Yoko Univ.of Tokyo, Grad.School of Arts and Sci., Assoc.Prof., 大学院・総合文化研究化, 助教授 (40158043)
• Co-Investigator(Kenkyū-buntansha): EDAMATSU Masaki Univ.of Tokyo, Grad.School of Arts and Sci., Res.Assistant, 大学院・総合文化研究化, 助手 (60251328)
• Project Period (FY): 1996 – 1997
• Project Status: Completed (Fiscal Year 1997)
• Budget Amount: ¥7,200,000 (Direct Cost: ¥7,200,000); Fiscal Year 1997: ¥1,800,000 (Direct Cost: ¥1,800,000); Fiscal Year 1996: ¥5,400,000 (Direct Cost: ¥5,400,000)
• Keywords: motor proteins / microtubules / kinesin / ncd / motor domain / neck region / polarity of the movement / 頸部 / ミオシン / 回転自由度

Research Abstract

Ncd, a member of kinesin super family, moves toward the minus end of microtubule. Neck region of kineshin and ncd is considered to be important to determine the polarity of the movement, because the structural difference between the motor domains of kineshin and ncd is not essential. We made a chymeric protein which consists of kinesin motor domain and ncd neck domain. This chymeric protein did not show the directional movement on microtubules. The measurement of ATPase activity revealed that the values of Kcat and K_<50%MT> are 7.8 S^<-1> and 0.22 muM,respectively. Comparing these values with those of kineshin fragment (kineshin motor domain and kineshin neck domain), the chymeric protein seemed to be unable to perform the processive movement. These results indicate that the neck region play an important role to determine the polarity of the movement.

Research Products

• [Publications] Inoue, Y., Toyodhima, Y.Y., Iwano, A, H., Morimoto, S., Higuchi, H., Yanagida, T.: "Movements of truncated kinesin fragments with a short or anartificial flexible neck" Proc.Natl.Acad.Sci.USA. 94. 7275-7280 (1997)
  • Description: 「研究成果報告書概要(和文)」より
• [Publications] Imafuku, Y., Toyoshima, Y.Y., &Tawda, K.: "Length dependence of displacement fluctuation and velocity in microtubule sliding movement driven by sea urchin sperm outer arm β dynein in vitro" Biophys.Chem.67. 117-125 (1997)
  • Description: 「研究成果報告書概要(和文)」より
• [Publications] 豊島 陽子: "微小管系モーターの多様な運動"
  • Description: 「研究成果報告書概要(和文)」より
• [Publications] Inoue, Y., Toyoshima, Y.Y., Iwane, A.H., Morimoto, S., Higuchi, H., & Yanagida, T.: "Movements of truncated kinesin fragments with a short or an artificial flexible neck" Proc.Natl.Acad.Sci.USA. 94. 7275-7280 (1997)
  • Description: 「研究成果報告書概要(欧文)」より
• [Publications] Imafuku, Y., Toyoshima, Y.Y., & Tawda, K.: "Length dependence of displacement fluctuations and velocity in microtubule sliding movement driven by sea urchin sperm outer arm beta dynein in vitro" Biophys.Chem.67. 117-125 (1997)
  • Description: 「研究成果報告書概要(欧文)」より
• [Publications] Lnoue,Y., Toyoshima,Y.Y., Iwane,A.H., Morimoto,S., Higuchi,H., Yanagida,T.: "Movements of truncated kinesin fragments with a short or an artificial flexible neck" Proc.Natl.Acad.Sci.USA. 94. 7275-7280 (1997)
• [Publications] Imafuku,Y., Toyoshima,Y.Y., & Tawda,K.: "Lenght dependence of displacement fluctuations and velocity in microtubule sliding movement driven by sea urchin sperm outer arm β dynein in vitro" Biophys.Chem.67. 117-125 (1997)
• [Publications] 豊島陽子: "細胞の中の動きを取り出して見る" 細胞のミクロ探検(日本学術協力財団). 15-25 (1997)
• [Publications] 豊島陽子: "In vitro motility assay" 細胞生物学の基礎技術-細胞骨格・細胞運動へのアプローチ(羊土社). 85-92 (1997)
• [Publications] 豊島陽子: "微小管系モーターの多様な運動" 生体分子モーターのしくみ(共立出版). 47-60 (1997)
• [Publications] Imafuku,Y.,Toyoshima,Y.& Tawada,K.: "A Monte Carlo study for a fluctuation analysis in the in vitro motility by protein motors." Biophys.Chem.59. 139-153 (1996)
• [Publications] Imafuku,Y.,Toyoshima,Y.& Tawada,K.: "Fluctuation in the microtubule sliding movement drinven by kinesin in vitro." Biophys.J.70. 878-886 (1996)
• [Publications] Tokai,N.,Fujimoto-Nishiyama,A.,Toyoshima,Y.Y.: "Kid,a novel kinesin-like DNA binding protein,is licalized to chromosomes and the mitotic spindle." The EMBO J.15. 457-467 (1996)
• [Publications] Edamatsu,M.,& Toyoshima,Y.Y.: "Isolation and characterization of pos mutants defective in correct positioning of septum in Schizosaccharomyces pombe." Zool.Sci.13. 235-239 (1996)