TY  - JOUR
AU  - Arakawa, M.
AU  - Yanamala, N.
AU  - Upadhyaya, J.
AU  - Halayako, A.
AU  - Klein-Seetharaman, J.
AU  - Chelikani, P.
TI  - The importance of valine 114 in ligand binding in beta(2)-adrenergic receptor
JO  - Protein science
VL  - 19
SN  - 0961-8368
CY  - Hoboken, NJ
PB  - Wiley
M1  - PreJuSER-8401
SP  - 85 - 93
PY  - 2010
N1  - Grant sponsor: Manitoba Medical Service Foundation (MMSF); Grant sponsor: Canadian Institute of Health Research; Grant number: ROP 92389; Grant sponsor: Manitoba Health Research Council (MHRC); Grant sponsor: National Institutes of Health; Grant number: NIH-NLM 2RO1LM007994-05; Grant sponsor: National Science Foundation; Grant number: CAREER NSF-CC0449117; Grant sponsor: New Investigator Personnel Award from the Heart and Stroke Foundation of Canada (HSFC).
AB  - G-protein coupled receptors (GPCRs) are transmembrane signaling molecules, with a majority of them performing important physiological roles. beta(2)-Adrenergic receptor (beta(2)-AR) is a well-studied GPCRs that mediates natural responses to the hormones adrenaline and noradrenaline. Analysis of the ligand-binding region of beta(2)-AR using the recently solved high-resolution crystal structures revealed a number of highly conserved amino acids that might be involved in ligand binding. However, detailed structure-function studies on some of these residues have not been performed, and their role in ligand binding remains to be elucidated. In this study, we have investigated the structural and functional role of a highly conserved residue valine 114, in hamster beta(2)-AR by site-directed mutagenesis. We replaced V114 in hamster beta(2)-AR with a number of amino acid residues carrying different functional groups. In addition to the complementary substitutions V114I and V114L, the V114C and V114E mutants also showed significant ligand binding and agonist dependent G-protein activation. However, the V114G, V114T, V114S, and V114W mutants failed to bind ligand in a specific manner. Molecular modeling studies were conducted to interpret these results in structural terms. We propose that the replacement of V114 influences not only the interaction of the ethanolamine side-chains but also the aryl-ring of the ligands tested. Results from this study show that the size and orientation of the hydrophobic residue at position V114 in beta(2)-AR affect binding of both agonists and antagonists, but it does not influence the receptor expression or folding.
KW  - Adenylate Cyclase: metabolism
KW  - Adrenergic beta-Antagonists: pharmacology
KW  - Alprenolol: pharmacology
KW  - Animals
KW  - Binding Sites
KW  - COS Cells
KW  - Cercopithecus aethiops
KW  - Cricetinae
KW  - Cyclic AMP: metabolism
KW  - Models, Molecular
KW  - Mutagenesis, Site-Directed
KW  - Protein Binding: drug effects
KW  - Receptors, Adrenergic, beta-2: chemistry
KW  - Receptors, Adrenergic, beta-2: genetics
KW  - Receptors, Adrenergic, beta-2: metabolism
KW  - Valine: chemistry
KW  - Valine: metabolism
KW  - Adrenergic beta-Antagonists (NLM Chemicals)
KW  - Receptors, Adrenergic, beta-2 (NLM Chemicals)
KW  - Alprenolol (NLM Chemicals)
KW  - Cyclic AMP (NLM Chemicals)
KW  - Valine (NLM Chemicals)
KW  - Adenylate Cyclase (NLM Chemicals)
KW  - J (WoSType)
LB  - PUB:(DE-HGF)16
C6  - pmid:19916165
C2  - pmc:PMC2817842
UR  - <Go to ISI:>//WOS:000273638700010
DO  - DOI:10.1002/pro.285
UR  - https://juser.fz-juelich.de/record/8401
ER  -