Arg 901 in the AE1 C-terminal tail is involved in conformational change but not in substrate binding

Biochim Biophys Acta. 2012 Mar;1818(3):658-65. doi: 10.1016/j.bbamem.2011.11.019. Epub 2011 Dec 1.

Abstract

In our previous paper, we demonstrated that Arg 901 in the C-terminal tail of human AE1 (band 3, anion exchanger 1) had a functional role in conformational change during anion exchange. To further examine how Arg 901 is involved in conformational change, we expressed various Arg 901 mutants and alanine mutants of the C-terminal tail (from Leu 886 to Val 911) on the plasma membrane of Saccharomyces cerevisiae and evaluated the kinetic parameters of sulfate ion transport. As a result, Vmax decreased as the hydrophobicities of the 901st and peripheral hydrophilic residues increased, indicating that the hydrophobicity of the C-terminal residue is involved in the conformational change. We also found the alkali and protease resistance of the C-terminal region after Arg 901 modification with hydroxyphenylglyoxal (HPG) or phenylglyoxal (PG), a hydrophobic reagent. These results suggested that the increased hydrophobicity of the C-terminal region around Arg 901 leads to inefficient conformational change by the newly produced hydrophobic interaction.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Anion Exchange Protein 1, Erythrocyte / chemistry
  • Anion Exchange Protein 1, Erythrocyte / genetics
  • Anion Exchange Protein 1, Erythrocyte / metabolism*
  • Arginine / chemistry
  • Arginine / genetics
  • Arginine / metabolism*
  • Cell Membrane / chemistry
  • Cell Membrane / genetics
  • Cell Membrane / metabolism*
  • Gene Expression
  • Humans
  • Hydrophobic and Hydrophilic Interactions
  • Ion Transport / physiology
  • Kinetics
  • Protein Binding / physiology
  • Protein Structure, Tertiary
  • Recombinant Proteins / genetics
  • Recombinant Proteins / metabolism
  • Saccharomyces cerevisiae

Substances

  • Anion Exchange Protein 1, Erythrocyte
  • Recombinant Proteins
  • Arginine