Isozymes of cAMP-dependent protein kinase present in the rat corpus luteum

M. Hunzicker-Dunn*, R. E. Cutler, E. T. Maizels, D. A. DeManno, M. L.G. Lamm, J. Erlichman, B. D. Sanwal, A. R. LaBarbera

*Corresponding author for this work

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27 Scopus citations

Abstract

Regulatory (R) subunits and their association with catalytic subunits to form cAMP-dependent protein kinase holoenzymes were investigated in corpora lutea of pregnant rats. Following separation by DEAE-cellulose chromatography, R subunits were identified by labeling with 8-N3[32P]cAMP and autophosphorylation on one and two-dimensional gel electrophoresis and by reactivity with antisera. DEAE-cellulose elution of R subunits with catalytic subunits as holoenzymes or without catalytic subunits was determined by sedimentation characteristics on sucrose density gradient centrifugation and by cAMP-stimulated kinase activation characteristics on Eadie-Scatchard analysis. We identified the presence of a type I holoenzyme containing RIα (M(r) 47,000) subunits, a prominent type II holoenzyme containing RIIβ (M(r) 52,000) subunits, and a second more acidic type II holoenzyme peak containing both RIIβ and RIIα (M(r) 54,000) subunits. However, the majority of total R subunit activity was associated with a catalytic subunit-free peak of RIα protein which on elution from DEAE-cellulose was associated with cAMP. This report establishes the more basic elution position from DEAE-cellulose of the prominent rat luteal RIIβ holoenzyme in very close proximity to free RIα and presents one of the few reports of a normal tissue containing a large percentage of catalytic subunit-free RIα.

Original languageEnglish (US)
Pages (from-to)7166-7175
Number of pages10
JournalJournal of Biological Chemistry
Volume266
Issue number11
StatePublished - 1991

ASJC Scopus subject areas

  • Biochemistry
  • Molecular Biology
  • Cell Biology

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    Hunzicker-Dunn, M., Cutler, R. E., Maizels, E. T., DeManno, D. A., Lamm, M. L. G., Erlichman, J., Sanwal, B. D., & LaBarbera, A. R. (1991). Isozymes of cAMP-dependent protein kinase present in the rat corpus luteum. Journal of Biological Chemistry, 266(11), 7166-7175.