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Prostaglandin E''-induced activation of adenosine 3'-5' cyclic monophosphate-dependent protein kinase of a murine macrophage-like cell line (P388D/sub 1/)

Journal Article · · J. Immunol.; (United States)
OSTI ID:5443941
;

Changes in the activities of adenosine 3',5'-cyclic monophosphate (cAMP)-dependent protein kinases in response to prostaglandin (PG)E/sub 2/-induced elevation of intracellular cAMP level were investigated with a murine macrophage-like cell line, P388D/sub 1/. Photoaffinity labeling with 8-azido-(/sup 32/P)cAMP showed that untreated P388D/sub 1/ cells possess two types of cAMP-binding proteins of m.w. 49,000 and 52,000, respectively, in the cytosol fraction in a ratio of 1:8. They must represent regulatory subunits (RI and RII, respectively) of cAMP-dependent protein kinases. Photoaffinity labeling of these fractions with 8-azido-(/sup 32/P)cAMP confirmed the separation of two types of isoenzymes, because each cAMP-dependent protein kinase active fraction was associated with only one type of regulatory subunit. The exposure of P388D/sub 1/ cells to exogenously added PGE/sub 2/ (1 ..mu..M) caused about 7.5-fold increase in the intracellular cAMP level within 30 sec. The enzyme assay of the cytosol demonstrated that the activation of cAMP-dependent protein kinases closely follows the kinetics of the intracellular cAMP level, which was measured by radioimmunoassay. The PGE/sub 2/-induced increase in the intracellular cAMP level appeared to activate preferentially the type I isoenzyme, inasmuch as the enzymatic activity of this type separated by the affinity chromatography of the cytosol of PGE/sub 2/-exposed cells was lower in the presence than in the absence of cAMP, whereas the type II enzyme activity remained responsive to exogenously added cAMP.

Research Organization:
Univ. of Kansas Medical Center, Kansas City
OSTI ID:
5443941
Journal Information:
J. Immunol.; (United States), Journal Name: J. Immunol.; (United States) Vol. 139:10; ISSN JOIMA
Country of Publication:
United States
Language:
English

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Related Subjects

550201 -- Biochemistry-- Tracer Techniques
550601* -- Medicine-- Unsealed Radionuclides in Diagnostics
59 BASIC BIOLOGICAL SCIENCES
62 RADIOLOGY AND NUCLEAR MEDICINE
AMP
ANIMAL CELLS
ANIMALS
BETA DECAY RADIOISOTOPES
BETA-MINUS DECAY RADIOISOTOPES
BIOLOGICAL EFFECTS
CONNECTIVE TISSUE CELLS
DAYS LIVING RADIOISOTOPES
ENZYME ACTIVITY
ENZYME INDUCTION
ENZYMES
GENE REGULATION
IMMUNOASSAY
IMMUNOLOGY
ISOTOPE APPLICATIONS
ISOTOPES
LABELLED COMPOUNDS
LIGHT NUCLEI
MACROPHAGES
MAMMALS
MICE
MOLECULAR STRUCTURE
NUCLEI
NUCLEOTIDES
ODD-ODD NUCLEI
ORGANIC COMPOUNDS
PHAGOCYTES
PHOSPHORUS 32
PHOSPHORUS ISOTOPES
PHOSPHORUS-GROUP TRANSFERASES
PHOSPHOTRANSFERASES
PROSTAGLANDINS
RADIOASSAY
RADIOIMMUNOASSAY
RADIOIMMUNOLOGY
RADIOISOTOPES
RODENTS
SOMATIC CELLS
TRACER TECHNIQUES
TRANSFERASES
TRITIUM COMPOUNDS
VERTEBRATES