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PMID: 8521811 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

Mutation of tyrosine-141 inhibits insulin-promoted tyrosine phosphorylation and increased responsiveness of the human beta 2-adrenergic receptor.

The EMBO journal ·Vol. 14 ·No. 22 ·1995-11-15 ·Pages 5542-9

Valiquette M, Parent S, Loisel TP, Bouvier M

Abstract

The ability of insulin to promote phosphorylation of the human beta 2-adrenergic receptor (beta 2AR) was assessed in Chinese hamster fibroblasts transfected with beta 2AR cDNA. Phosphotyrosine residues were detected in purified beta 2AR using a polyclonal anti-phosphotyrosine antibody and by phosphoamino acid analysis following metabolic labelling with inorganic 32P. Treatment of the cells with insulin induced a 2.4-fold increase in the phosphotyrosine content of the receptor. The insulin-promoted phosphorylation of the beta 2AR was accompanied by an increase in the beta-adrenergic-stimulated adenyl cyclase activity. Substitution of a phenylalanine residue for tyrosine-141 completely prevented both the increased tyrosine phosphorylation and the enhanced responsiveness of the beta 2AR promoted by insulin treatment. Mutation of three other tyrosines located in the cytoplasmic domain of the receptor, tyrosine-366, tyrosine-350 and tyrosine-354, did not abolish the insulin-promoted tyrosine phosphorylation. Taken together, these results suggest that insulin promotes phosphorylation of the beta 2AR on tyrosine-141 and that such phosphorylation leads to a supersensitization of the receptor.

MeSH Terms
Amino Acid Sequence Animals Base Sequence Binding Sites Cells, Cultured Cricetinae DNA Primers Fibroblasts/cytology Humans Insulin/pharmacology Molecular Sequence Data Mutagenesis, Site-Directed Phosphorylation Phosphotyrosine/metabolism Protein Conformation Receptors, Adrenergic, beta-2/chemistry,genetics,metabolism Structure-Activity Relationship Transfection Tyrosine/genetics,metabolism
Chemicals
DNA Primers Insulin Receptors, Adrenergic, beta-2 Phosphotyrosine Tyrosine
Authors & Affiliations
4 authors, click to expand affiliations / ORCID
Valiquette M
Département de Biochimie, Université de Montréal, Qc, Canada.
Parent S
Loisel T P
Bouvier M
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Article Info
Journal
The EMBO journal
Abbr.
EMBO J
ISSN
0261-4189
Published
1995-11-15
Pages
5542-9
Language
English
Region
England
NLM ID
8208664
PMCID
PMC394668
Subset
IM
External Links
PubMed source PMC Full Text View journal details
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