Identification of a Ca2+/calmodulin‐binding domain within the carboxyl‐terminus of the angiotensin II (AT1A) receptor
Open Access
- 23 July 1999
- journal article
- Published by Wiley in FEBS Letters
- Vol. 455 (3) , 367-371
Abstract
To identify regulators of the type 1A angiotensin II receptor (AT1A), we investigated the interaction of cellular proteins with a fusion protein containing the rat AT1A receptor carboxyl‐terminus. An ∼20 kDa cytoplasmic protein interacted with the fusion protein in a Ca2+‐dependent manner and was identified as calmodulin. A control peptide with high affinity for Ca2+/calmodulin and a peptide corresponding to a membrane proximal portion of the AT1A receptor carboxyl‐terminus with analogy to known calmodulin‐binding sequences were synthesised and tested for calmodulin‐binding. Using in vitro binding assays combined with gel shift analysis, we demonstrated the formation of complexes between calmodulin and both peptides, which were Ca2+‐dependent and of 1:1 stoichiometry. Affinity gels produced from these peptides also purified calmodulin from cell extracts. These results suggest a novel feedback regulation of the AT1A receptor by Ca2+/calmodulin and identify the membrane proximal region of the carboxyl‐terminus as a focal point for interactions important for AT1A receptor function.Keywords
This publication has 35 references indexed in Scilit:
- Molecular tinkering of G protein‐coupled receptors: an evolutionary successThe EMBO Journal, 1999
- Angiotensin II-induced Association of Phospholipase Cγ1 with the G-protein-coupled AT1 ReceptorJournal of Biological Chemistry, 1998
- Dependence on the Motif YIPP for the Physical Association of Jak2 Kinase with the Intracellular Carboxyl Tail of the Angiotensin II AT1 ReceptorJournal of Biological Chemistry, 1997
- Phosphorylation and Calmodulin Binding of the Metabotropic Glutamate Receptor Subtype 5 (mGluR5) Are Antagonistic in VitroJournal of Biological Chemistry, 1997
- Calcium/Calmodulin‐Dependent Kinase II Phosphorylates Drosophila Visual ArrestinJournal of Neurochemistry, 1997
- Identification of an Essential Signaling Cascade for Mitogen-activated Protein Kinase Activation by Angiotensin II in Cultured Rat Vascular Smooth Muscle CellsJournal of Biological Chemistry, 1996
- Inactivation of NMDA Receptors by Direct Interaction of Calmodulin with the NR1 SubunitCell, 1996
- The role of tyrosine phosphorylation in angiotensin II-mediated intracellular signallingCardiovascular Research, 1995
- Calmodulin-binding domains: just two faced or multi-faceted?Trends in Biochemical Sciences, 1995
- How calmodulin binds its targets: sequence independent recognition of amphiphilic α-helicesTrends in Biochemical Sciences, 1990