• Search Research Projects
  • Search Researchers
  • How to Use
  1. Back to project page

1988 Fiscal Year Final Research Report Summary

PROTEIN CHEMISTRY AND BIOCHEMICAL STUDY ON THE ANION TRANSPORT SYSTEM IN HUMAN ERYTHROCYTE MEMBRANES.

Research Project

Project/Area Number 61570149
Research Category

Grant-in-Aid for General Scientific Research (C)

Allocation TypeSingle-year Grants
Research Field Pathological medical chemistry
Research InstitutionFukuoka University

Principal Investigator

HAMASAKI Naotaka  DEPART. OF CLINICAL CHEMISTRY AND LAB. MED., FUKUOKA UNIV. SCHOOL OF MED., 医学部, 教授 (00091265)

Project Period (FY) 1986 – 1988
KeywordsAnion transport system / Erythrocyte membrane / Transmembrane protein / Amino acid sequence / Active center / Band 3
Research Abstract

A human erythrocyte band 3 peptide, affinity labeled with pyridoxal phosphate, was purified by a combination of gel permeation and reverse-phase high performance liquid chromatography. The amino acid sequence of the transmembrane peptide was determined by sequencing subfragments of the peptide obtained from lysyl endopeptidase and staphlococcal proteinase V8 digestions.
When a peptide containing the COOH-terminal of human erythrocyte Band 3 was also purified and sequenced, the affinity-labeled peptide was found to be located close to the COOH-terminal of Band 3, where it could be aligned with amino acid residues 852-927 of a murine erythrocyte Band 3, deduced from a nucleotide seguence of a cDNA clone. The amino acid sequence of the COOH-terminal region was highly homologous to that of murine Band 3.
As a result, the sequence of the COOH-terminal peptide of band 3 was established as follows. H^^1LFTGIQIIX^^<10> LAVLWVVKST^^<20> PASLALPFVL^^<30> ILTVPLRRVL^^<40> LPLIF RNVEL^^<50> QCLDADDAKA^^<60> TFDEEEGRDE^^<70> YDEVAMPV^^<78>
The pyridoxal phosphate binding site was identified as Lys-18 which corresponded to Lys-869 of the deduced sequence. It appears that the COOH-terminal region of band 3 constitutes at least a part of the active center for anion transport in human erythrocyte membrans.
Diethylpyrocarbonate inhibited the phosphate exchange across the human erythrocyte membrane. The exchange rate was inhibited only when the membranes were modified with the reagent from the cytosolic surface of resealed ghosts. The intracellular modification by diethylpyrocarbonate inhibited the extracellular binding of [^3H@]-DIDS to Band 3 protein. Furthermore, the extracellular DNDS protected the membranes from the intracellular modification by diethylpyrocarbonate.
The sequenced COOH-terminal peptide closely related to the DIDS binding site. Thus, the intracellular modification by diethylpyrocarbonate may transmembranously change the conformation of the COOH-terminal peptide.

  • Research Products

    (13 results)

All Other

All Publications (13 results)

  • [Publications] Kawano,Y.;Hamasaki,N.: J.Biochem.100. 191-199 (1986)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Hamasaki,N.;Kawano,Y.: Trends in Biochem.Sci(TIBS)(Review). 12. 183-185 (1987)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Inoue,H.;Moriyasu,M.;Hamasaki,N.: J.Biol.Chem.262. 7635-7638 (1987)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Kawano,Y.;Okubo,K.;Tokunaga,F.;Miyata,T.;Iwanaga,S.;Hamasaki,N.: J.Biol.Chem.263. 8232-8238 (1988)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Igisu,H.;Hamasaki,N.;Ito,A.;Ou,W.: Lipids. 23. 345-348 (1988)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Izuhara,K.;Hamasaki,N.: Biochemistry. (1989)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] 濱崎直孝: "赤血球膜蛋白質の構造と機能" 宇宙堂八木書店, 87 (1987)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] 河野洋一,濱崎直孝: "続生化学実験講座8血液(上)" 化学同人, 476 (1987)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Kawano, Y.; Hamasaki, N.: "Isolation of a 5,300-Dalton Peptide Containing a Pyridoxal Phosphate binding Site from the 38,000-Dalton of Band 3 of Human Erythrocyte Membrane." J. Biochem.100. 191-199 (1986)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Hamasaki, N.; Kawano, Y.: "Phosphoenolpyruvate Transport by the Anion Transport System of Human Erythrocyte Membranes." Trends in Biochem. Sci. (TIBS). 12. 183-185 (1987)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Inoue, H.; Moriyasu, M.; Hamasaki, N.: "Metabolism of 3-phosphoglyceroylphosphate in Phosphoenolpyruvate-enriched Human Erythrocytes." J. Biol. Chem.262. 7635-7638 (1987)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Kawano, Y.; Okubo, K.; Tokunaga, F.; Miyata, T.; Iwanaga, S.; Hamasaki, N.: "Localization of the Pyridoxal Phosphate Binding Site at the COOH-terminal Region of Erythrocyte Band 3 Protein." J. Biol. Chem.263. 8232-8238 (1988)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Igisu, H.; Hamasaki, N.; Ito, A.; Ou,W.: "Inhibition of Cytochrome c Oxidase and Hemolysis Caused by Lysosphingolipids." Lipids. 23. 345-348 (1988)

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

URL: 

Published: 1990-03-20  

Information User Guide FAQ News Terms of Use Attribution of KAKENHI

Powered by NII kakenhi