Sun H, Seyer J M, Patel T B
Department of Pharmacology, University of Tennessee, Memphis 38163.
Proc Natl Acad Sci U S A. 1995 Mar 14;92(6):2229-33. doi: 10.1073/pnas.92.6.2229.
Epidermal growth factor (EGF) stimulates adenylyl cyclase in the heart via activation of the stimulatory GTP-binding protein Gs. Therefore, employing peptides corresponding to regions in the cytosolic domain of the EGF receptor, we have investigated the ability of sequences within the EGF receptor to activate Gs. A 13-aa peptide (EGFR-13) corresponding to the juxtamembrane region in the cytosolic domain of the EGF receptor stimulated GTP binding and GTPase activity of Gs. This peptide did not stimulate GTP binding to Gi but increased the GTPase activity of this protein. Additionally, phosphorylation of the protein kinase C site (threonine residue) within EGFR-13 decreased the ability of the peptide to stimulate Gs and increase GTPase activity of Gi. Further, in functional assays of Gs employing S49 cyc- cell membranes, EGFR-13 increased the ability of Gs to stimulate adenylyl cyclase; phospho-EGFR-13 and a 14-aa peptide corresponding to a sequence in the cytosolic domain of the EGF receptor did not alter the functional activity of Gs. Hence, the juxtamembrane region of the EGF receptor can activate Gs and, by stimulating GTPase activity of Gi, inactivates this latter G protein. Phosphorylation of the threonine residue within this region attenuates the activity of the peptide as a modulator of G-protein function.
表皮生长因子(EGF)通过激活刺激性GTP结合蛋白Gs来刺激心脏中的腺苷酸环化酶。因此,我们利用与EGF受体胞质结构域区域相对应的肽段,研究了EGF受体中的序列激活Gs的能力。一种与EGF受体胞质结构域中近膜区域相对应的13个氨基酸的肽段(EGFR - 13)刺激了Gs的GTP结合和GTP酶活性。该肽段不刺激GTP与Gi的结合,但增加了该蛋白的GTP酶活性。此外,EGFR - 13内蛋白激酶C位点(苏氨酸残基)的磷酸化降低了该肽段刺激Gs和增加Gi的GTP酶活性的能力。此外,在使用S49 cyc - 细胞膜对Gs进行的功能测定中,EGFR - 13增强了Gs刺激腺苷酸环化酶的能力;磷酸化的EGFR - 13和一种与EGF受体胞质结构域序列相对应的14个氨基酸的肽段并未改变Gs的功能活性。因此,EGF受体的近膜区域可以激活Gs,并通过刺激Gi的GTP酶活性来使后者失活。该区域内苏氨酸残基的磷酸化减弱了该肽段作为G蛋白功能调节剂的活性。