Angiotensin II type 1 receptor-function affected by mutations in cytoplasmic loop CD

FEBS Lett. 2000 Mar 31;470(3):331-5. doi: 10.1016/s0014-5793(00)01346-6.

Abstract

To explore peptide hormone-induced conformational changes, we attempted to engineer a metal-ion binding site between the cytoplasmic loops CD and EF in the angiotensin II type 1 (AT(1)) receptor. We constructed 12 double and six triple histidine mutant receptors, and tested the ability of each mutant and the wild-type to activate inositol phosphate (IP) production with and without ZnCl(2). Inhibition by ZnCl(2) in the double and triple His mutant receptors was not significant, but these mutations directly decreased the IP production. Systematic analysis of single His mutants demonstrated that the loop CD-mutants displayed 52-74% inhibition of IP production, whereas the loop EF-mutants did not affect IP production. These results indicate that the cytoplasmic loop CD-segment from Tyr(127) to Ile(130) is important for G(q/11) activation by the AT(1) receptor.

Publication types

  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Amino Acid Sequence
  • Amino Acid Substitution / genetics
  • Angiotensin II / metabolism
  • Angiotensin II / pharmacology
  • Angiotensin Receptor Antagonists
  • Animals
  • Binding Sites
  • COS Cells
  • Cattle
  • Chlorides / metabolism
  • Chlorides / pharmacology
  • Heterotrimeric GTP-Binding Proteins / metabolism
  • Histidine / genetics
  • Inositol Phosphates / metabolism
  • Kinetics
  • Losartan / metabolism
  • Losartan / pharmacology
  • Models, Molecular
  • Molecular Sequence Data
  • Mutation / genetics*
  • Protein Structure, Secondary
  • Receptor, Angiotensin, Type 1
  • Receptor, Angiotensin, Type 2
  • Receptors, Angiotensin / chemistry*
  • Receptors, Angiotensin / genetics
  • Receptors, Angiotensin / metabolism*
  • Rhodopsin / chemistry
  • Sequence Alignment
  • Signal Transduction / drug effects
  • Thermodynamics
  • Zinc Compounds / metabolism
  • Zinc Compounds / pharmacology

Substances

  • Angiotensin Receptor Antagonists
  • Chlorides
  • Inositol Phosphates
  • Receptor, Angiotensin, Type 1
  • Receptor, Angiotensin, Type 2
  • Receptors, Angiotensin
  • Zinc Compounds
  • Angiotensin II
  • Histidine
  • zinc chloride
  • Rhodopsin
  • Heterotrimeric GTP-Binding Proteins
  • Losartan