Immunological distinction between guanosine 3':5'-monophosphate-dependent and adenosine 3':5'-monophosphate-dependent protein kinases.

U Walter,P Miller,F Wilson,D Menkes,P Greengard

doi:10.1016/s0021-9258(19)85769-1

Abstract

A guanosine 3':5'-monophosphate (cGMP)-dependent protein kinase was purified from bovine lung using 8-(6-aminohexylamino)-cAMP-Sepharose. The activity of the purified enzyme was highly dependent on cGMP using histone f2b as a substrate. The self-phosphorylation of the purified enzyme was strongly inhibited by cGMP and not significantly affected by cAMP. A precipitating antiserum prepared in rabbits against the cGMP-dependent protein kinase specifically inhibited the histone kinase activity and the self-phosphorylation of the purified cGMP-dependent protein kinase without affecting the cGMP binding site. This antiserum also specifically inhibited the phosphorylation of the endogenous substrate proteins by endogenous cGMP-dependent protein kinase in smooth muscle membranes, but did not cross-react detectably with catalytic subunit or regulatory subunit of type I or type II cAMP-dependent protein kinase. Conversely, anti-sera against the regulatory subunit of type I or type II cAMP-dependent protein kinase did not cross-react detectably with cGMP-dependent protein kinase. The substantial differences between the immunological properties of the cGMP-dependent and cAMP-dependent protein kinases suggest that these two enzymes have distinct physiological roles.

Highlights

(6-aminohexylamino)-cAMP-Sepharose.The activity of ever, specific endogenous substrates for the cGMP-dependent thepurifiedenzyme was highly dependent on cGMP protein kinase have been described (12-X), and cGMP-deusing histone fibas a substrate
The elution of the cCMP-dependent protein kinase pared from bovine pulmonary arteries as described [20].Type I and from the AHA-CAMP-Sepharose column was continued as de- type I1 regulatory subunits (R-I andR-11) and the cGMP-dependent scribed in detail in the legend of Fig. l
It was noted that the presence of 10 pl of control serum slightly stimulated the activity of the cGMP-dependent protein kinase and slightly inhibited the activity of the catalytic subunit of CAMP-dependentprotein kinase

Summary

Introduction

(6-aminohexylamino)-cAMP-Sepharose.The activity of ever, specific endogenous substrates for the cGMP-dependent thepurifiedenzyme was highly dependent on cGMP protein kinase have been described (12-X), and cGMP-deusing histone fibas a substrate. The elution of the cCMP-dependent protein kinase pared from bovine pulmonary arteries as described [20].Type I and from the AHA-CAMP-Sepharose column was continued as de- type I1 regulatory subunits (R-I andR-11) and the cGMP-dependent scribed in detail in the legend of Fig. l.

Results

Conclusion