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Characterization of cell surface adenosine 3',5'-monophosphate-binding proteins in Y-1 mouse adrenal tumor cells

Journal Article · · Endocrinology; (United States)
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Adrenal cortical cells are known to export cAMP and have binding proteins and cAMP-dependent protein kinase activity associated with their plasma membranes. Because these properties suggest a function for extracellular cAMP, we have undertaken a search for specific cell surface receptors for this cyclic nucleotide. Y-1 mouse adrenal tumor cells actively export cAMP by an energy-dependent process. Analysis of Scatchard plots of the equilibrium binding of (/sup 3/H)cAMP to these cells indicate the existence of two classes of cAMP binders: one with high affinity (K/sub a/ . 2.9 X 10(9) M-1) and another with low affinity (K/sub a/ . 7.0 X 10(7) M-1). The cell surface localization of these binders was established by the sensitivity of both the (/sup 3/H)cAMP-binding proteins and the (/sup 32/P)8-N3-cAMP photoaffinity labeled proteins of intact cells to mild trypsin digestion and by the surface distribution of a BSA-O2-monosuccinyl cAMP-gold complex revealed by electron microscopy. Analysis of radioautograms of cell surface cAMP-binding proteins from confluent monolayer tumor cells, photoaffinity labeled with (/sup 32/P)8-N3-cAMP and subjected to sodium dodecyl sulfate-polyacrylamide gel electrophoresis revealed two major /sup 32/P-labeled protein bands which were indistinguishable from the 49,000 and 55,000 mol wt regulatory subunits of the cytosolic protein kinase isoenzymes of this cell. These observations along with the demonstration of cell surface, cAMP-dependent protein kinase activity in the mouse adrenal tumor cell strongly suggest that these cAMP-binding proteins function as regulatory proteins for cell surface protein kinases.

Research Organization:
Univ. of Southern California, Los Angeles
OSTI ID:
5483466
Journal Information:
Endocrinology; (United States), Journal Name: Endocrinology; (United States) Vol. 116:3; ISSN ENDOA
Country of Publication:
United States
Language:
English

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Related Subjects

550201 -- Biochemistry-- Tracer Techniques
550501* -- Metabolism-- Tracer Techniques
59 BASIC BIOLOGICAL SCIENCES
ADRENAL GLANDS
AMP
ANIMALS
AUTORADIOGRAPHY
BETA DECAY RADIOISOTOPES
BETA-MINUS DECAY RADIOISOTOPES
BIOCHEMICAL REACTION KINETICS
BODY
CELL CULTURES
DAYS LIVING RADIOISOTOPES
DISEASES
DISTRIBUTION
ELECTRON MICROSCOPY
ELECTROPHORESIS
ENDOCRINE GLANDS
ENZYME ACTIVITY
ENZYMES
GLANDS
ISOTOPE APPLICATIONS
ISOTOPES
KINETICS
LABELLED COMPOUNDS
LIGHT NUCLEI
MAMMALS
MEMBRANE TRANSPORT
MICE
MICROSCOPY
NEOPLASMS
NUCLEI
NUCLEOTIDES
ODD-ODD NUCLEI
ORGANIC COMPOUNDS
ORGANS
PHOSPHORUS 32
PHOSPHORUS ISOTOPES
PHOSPHORUS-GROUP TRANSFERASES
PHOSPHOTRANSFERASES
RADIOISOTOPES
REACTION KINETICS
RECEPTORS
RODENTS
TISSUE DISTRIBUTION
TRACER TECHNIQUES
TRANSFERASES
TRITIUM COMPOUNDS
VERTEBRATES