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Effects of human erythrocyte guanine nucleotide-binding regulatory protein on parathyroid hormone-responsive adenylate cyclase from canine renal cortex.

Abstract：

:We studied the effects of the guanine nucleotide-binding regulatory protein (Gs) from human erythrocytes on PTH-responsive adenylate cyclase from partially purified membranes of canine renal cortex (CRC). Extracts of erythrocyte membranes, containing soluble Gs, was obtained by treatment with a detergent (Lubrol PX). Gs did not stimulate adenylate cyclase activity by itself, but amplified the response of adenylate cyclase in CRC membranes to both synthetic bovine PTH-(1-34) [bPTH-(1-34)] and to the hydrolysis-resistant GTP analog 5'-guanylimido-diphosphate [Gpp(NH)p]. Gs increased PTH stimulation of adenylate cyclase activity in both the presence and absence of Gpp(NH)p. In the absence of Gpp(NH)p, the potentiating effect of Gs occurred only when the concentration of bPTH-(1-34) was greater than 10 ng/ml. bPTH-(1-34), Gpp(NH)p, and Gs each enhanced the catalytic activity of adenylate cyclase when added separately or in combination by increasing the apparent maximum velocity (Vmax) of the enzyme without altering the apparent Km for MgATP. The effect of Gs on CRC membrane adenylate cyclase activity in the presence of NaF (10 mM) and forskolin (100 microM) was also examined. NaF- and forskolin-stimulated enzyme activities were significantly increased by Gs in both the presence and absence of Gpp(NH)p (100 microM). Analysis of double reciprocal plots of substrate concentration and enzyme activity revealed that NaF and forskolin increased the Vmax of the catalytic activity and did not alter the apparent Km of the enzyme for MgATP. These data support the role of Gs as a regulator of the response of adenylate cyclase to hormones, guanyl nucleotides, NaF, and forskolin. Our studies address the relative functional stoichiometry between Gs and catalytic unit present in CRC membranes and suggest that the CRC adenylate cyclase system must contain insufficient Gs to couple with all available catalytic units. These results are consistent with the possibility that deficiency of Gs impairs hormonal stimulation by diminishing the apparent Vmax of the catalytic unit and does not alter the apparent affinity of the enzyme for MgATP.

journal_name

Endocrinology

journal_title

Endocrinology

authors

Levine MA,Greene A,Turner RT,Bell NH

doi

10.1210/endo-115-4-1386

subject

Has Abstract

pub_date

1984-10-01 00:00:00

pages

1386-91

issue

eissn

0013-7227

issn

1945-7170

journal_volume

115

pub_type

杂志文章

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Effects of human erythrocyte guanine nucleotide-binding regulatory protein on parathyroid hormone-responsive adenylate cyclase from canine renal cortex.