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Autophosphorylation and dephosphorylation of type II cAMP-dependent protein kinase in intact tissue and cells

Thesis/Dissertation ·
OSTI ID:6281614

Monoclonal antibodies were used to quantitate changes in the extent of autophosphorylation of the type II regulatory subunit of cAMP-dependent protein kinase in intact bovine tracheal smooth muscle and rat hepatocytes. The autophosphorylated and dephosphorylated forms of the regulatory subunit (RII) were separated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis, and identified by immunoblot analysis. Incubating smooth muscle strips with agents which stimulate cAMP production caused a rapid and reversible dephosphorylation of phospho RII. In contrast, phospho RII levels in rat hepatocytes were unaffected by elevations in cAMP concentration. Insulin was capable of inhibiting cAMP-dependent protein kinase in hepatocytes. Tissue extracts were tested to identify the basis for the lack of RII dephosphorylation in intact hepatocytes. Rat liver extract contained 4 fold less RII and had an 80 fold slower rate of dephosphorylation of endogenous RII compared to bovine smooth muscle extract. The different rates were not observed using purified, /sup 32/P-labelled RII and tissue extracts suggesting the decreased RII dephosphorylation rate in liver was due to a difference in the availability of endogenous RII rather than a difference in measurable phosphatase activity.

Research Organization:
Texas Univ., Dallas, TX (USA). Southwestern Medical Center
OSTI ID:
6281614
Country of Publication:
United States
Language:
English

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Related Subjects

550201* -- Biochemistry-- Tracer Techniques
59 BASIC BIOLOGICAL SCIENCES
AMP
ANIMAL CELLS
ANIMALS
ANTIBODIES
BIOSYNTHESIS
CATTLE
CHEMICAL REACTIONS
DOMESTIC ANIMALS
ELECTROPHORESIS
ENZYME ACTIVITY
ENZYMES
ISOTOPE APPLICATIONS
ISOTOPES
LIGHT NUCLEI
LIVER CELLS
MAMMALS
MONOCLONAL ANTIBODIES
MUSCLES
NUCLEI
NUCLEOTIDES
ODD-EVEN NUCLEI
ORGANIC COMPOUNDS
PHOSPHORUS 31
PHOSPHORUS ISOTOPES
PHOSPHORUS-GROUP TRANSFERASES
PHOSPHORYLATION
PHOSPHOTRANSFERASES
RATS
RESPIRATORY SYSTEM
RODENTS
RUMINANTS
SOMATIC CELLS
STABLE ISOTOPES
SYNTHESIS
TRACER TECHNIQUES
TRACHEA
TRANSFERASES
VERTEBRATES