The angiotensin II AT1 receptor is tyrosine and serine phosphorylated and can serve as a substrate for the src family of tyrosine kinases.
Angiotensin II AT1 receptor signal transduction has recently been shown to function through the phospholipase C isozyme, PLC-gamma. Since PLC-gamma is known to interact with phosphotyrosine containing proteins through SH2 domains, we examined the phosphorylation state of the AT1 receptor. Immunoprecipitation of the [32P] labeled AT1 receptor from rat aortic smooth muscle cells followed by alkali hydrolysis demonstrated the presence of tyrosine phosphorylation. Phosphoamino acid analysis of the excised bands demonstrated the presence of phosphoserine and phosphotyrosine residues. A fusion protein comprising the intracellular tail of the AT1 receptor was used to screen for candidate kinases, and the src kinase family displayed high activity. In summary, this study shows that the AT1 receptor is serine and tyrosine phosphorylated in vivo and suggests that a soluble kinase related to the src family may be responsible for the tyrosine phosphorylation.
Paxton
W G
WG
Department of Pathology, Emory University, Atlanta, GA 30322.
Marrero
M B
MB
Klein
J D
JD
Delafontaine
P
P
Berk
B C
BC
Bernstein
K E
KE
D000804:Angiotensin II / Q000378:metabolism*; D000818:Animals; D001011:Aorta / Q000378:metabolism; D002478:Cells, Cultured; D003001:Cloning, Molecular; D016391:Genes, src; D005982:Glutathione Transferase / Q000096:biosynthesis / Q000378:metabolism; D007527:Isoenzymes / Q000378:metabolism; D007700:Kinetics; D009131:Muscle, Smooth, Vascular / Q000378:metabolism*; D010710:Phosphates / Q000378:metabolism; D010761:Phosphorus Radioisotopes; D010766:Phosphorylation; D010768:Phosphoserine / Q000032:analysis*; D019000:Phosphotyrosine; D051381:Rats; D020794:Receptor Protein-Tyrosine Kinases / Q000378:metabolism*; D011945:Receptors, Angiotensin / Q000096:biosynthesis / Q000302:isolation & purification / Q000378:metabolism*; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis / Q000302:isolation & purification / Q000378:metabolism; D013379:Substrate Specificity; D010738:Type C Phospholipases / Q000378:metabolism; D014443:Tyrosine / Q000031:analogs & derivatives* / Q000032:analysis
D007527:Isoenzymes; D010710:Phosphates; D010761:Phosphorus Radioisotopes; D011945:Receptors, Angiotensin; D011993:Recombinant Fusion Proteins; D000804:Angiotensin II; D010768:Phosphoserine; D019000:Phosphotyrosine; D014443:Tyrosine; D005982:Glutathione Transferase; D020794:Receptor Protein-Tyrosine Kinases; D010738:Type C Phospholipases