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2002 Fiscal Year Final Research Report Summary

Structure and function of ion-channel VO of Na+-translocating V-ATPase

Research Project

Project/Area Number 13672272
Research Category

Grant-in-Aid for Scientific Research (C)

Allocation TypeSingle-year Grants
Section一般
Research Field Biological pharmacy
Research InstitutionMURORAN INSTITUTE OF TECHNOLOGY

Principal Investigator

KAKINUMA Yoshimi  Muroran Institute of Technology, Faculty of Engineering, Professor, 工学部, 教授 (80134394)

Project Period (FY) 2001 – 2002
KeywordsV-ATPase / Sodium / Ion / Enterococcus hirae
Research Abstract

The role of NtpI C-terminal region of E. hirae Na^+-ATPase was studied. In this region, there is an arginine residue, which is highly conserved among the corresponding subunits of bacterial V-ATPases, at position 573 of NtpI. Substitution of Arg-573 with Glu, Leu, or Gln abolished sodium transport and sodium-stimulated ATP hydrolysis of the enzyme. The conservative replacement of Arg by Lys lowered both activities to about one fifth of those of the wild-type enzyme. We previously reported an ATP-dependent negative cooperativity for Na^+ coupling of this enzyme. This negative cooperativity was weakened by the mutation Arg573Lys; the Hill coefficients for the wild type and mutant enzymes at a saturated ATP concentration were 0.22±0.03 and 0.40±0.05, respectively. The Hill coefficients of both enzymes at limited ATP concentrations approached 1. These results indicate that NtpI Arg573 is indispensable for sodium translocation and for the cooperative features of E. hirae V-ATPase.
In addition to Arg-573, the Tyr571, Leu574 and Leu577 were also involved in Na^+ translocation. And other important residues, His-626 and Glu-634, were also required for Na^+-ATPase activity, but their role in ion translocation may differ from conserved His or Glu residues of other a subunits. From these results, it is indicated that the C-terminal region of NtpI is both essential in sodium translocation and stabilization of NtpI.

  • Research Products

    (16 results)

All Other

All Publications (16 results)

  • [Publications] Murata T. et al.: "Nucleotide-Binding Sites in V-Type Na+-ATPase from Enterococcus hirae"Journal of Biochemistry (Tokyo). 132. 789-794 (2002)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Kawano, M. et al.: "Isolation of Enterococcus hirac mutant deficient in low-affinity potassium uptake at alkaline pH"Bioscience, Biotechnology and Biochemistry. 66. 1597-1600 (2002)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Yasumura, K. et al.: "Promoter analysis of the sodium-responsive V-ATPase (ntp) operon in Enterococcus hirae"Archives of Microbiology. 178. 172-179 (2002)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Kawano, M. et al.: "Arginine residue at position 573 in Enterococcus hirae vacuolar-type ATPase NtpI subunit plays a crucial role in Na+ translocation"Journal of Biological Chemistry. 277. 24405-24410 (2002)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Murata, T. et al.: "The membrane domain of the Na+ motive V-ATPase from Enterococcus hirae contains a heptameric rotor"Journal of Biological Chemistry. 278(印刷中). (2003)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Murata T. et al.: "Catalytic properties of Na+ -translocating V-ATPase in Enterococcus hirae"Biochim.Biophys.Acta. 1505. 75-81 (2001)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Kawano, M. et al.: "PotassiumuptakewithlowaffinityandhighrateinEnterococcushiraeatalkalinepH"Archives of Microbiology. 175. 41-45 (2001)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Murata, T. et al.: "ATP-dependent affinity change of Na+ binding sites of V-ATPase in Enterococcus hirae"Journal of Biological Chemistry. 276. 48337-48340 (2001)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Murata, T., Kakinuma, Y., Yamato, I.: "ATP-dependent affinity change of Na^+ binding sites of V-ATPase in Enterococcus hirae"J. Biol. Chem.. 276. 48337-48340 (2001)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Murata, T., Kawano, M., Igarashi, K., Yamato, I., Kakinuma, Y.: "Catalytic properties of Na^+-translocating V-ATPase in Enterococcus hirae"Biochim. Biophys. Acta (Bioenergetics). 1505. 75-81 (2001)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Kawano, M., Abuki, R., Igarashi, K., Kakinuma, Y.: "Potassium uptake with low affinity and high rate in Enterococcus hirae at alkaline pH"Arch. Microbiol.. 175. 41-45 (2001)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Kawano, M., Igarashi, K., Yamato, I., Kakinuma, Y.: "Arginine residue at position 573 in Enterococcus hirae V-ATPase NtpI subunit plays a crucial role in Na^+ translocation"J. Biol. Chem.. 277. 24405-24410 (2002)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Yasumura, K., Igarashi, K., Kakinuma, Y.: "Promoter analysis of the sodium-responsive V-ATPase (ntp) operon in Enterococcus hirae"Arch. Microbiol.. 178. 172-179 (2002)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Kawano, M., Igarashi, K., Kakinuma, Y.: "Isolation of Enterococcus hirae mutant deficient in low-affinity potassium uptake at alkaline pH"Biosci. Biotechnol. Biochem.. 66. 1597-1600 (2002)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Murata, T., Yoshikawa, Y., Hosaka, T., Takase, K., Kakinuma, Y., Yamato, I., Kikuchi, T.: "Nucleotide binding sites in V-type Na^+-ATPase from Enterococcus hirae"J. Biochem (Tokyo). 132. 789-794 (2002)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Murata, T., Arechaga, I., Fearnley, I. M., Kakinuma, Y., Yamato, I., Walker, J. E.: "The membrane domain of the Na^+-motive V-ATPase from Enterococcus hirae contains a heptameric rotor"J. Biol. Chem.. in press. (2003)

    • Description
      「研究成果報告書概要(欧文)」より

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Published: 2004-04-14  

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