Glucocorticoid regulation in rat brain cell cultures. Hydrocortisone increases the rate of synthesis of glycerol phosphate dehydrogenase in C6 glioma cells. [Tritium tracer technique]

Journal Article · Sun Dec 10 04:00:00 EST 1978 · J. Biol. Chem.; (United States)

OSTI ID:6803755

McGinnis, J F; de Vellis, J

Cytoplasmic glycerol phosphate dehydrogenase (sn-glycerol-3-phosphate: NAD/sup +/ 2-oxidoreductase, EC 1.1.1.8) was rapidly purified from rat skeletal muscle in high yield using a combination of classical and affinity techniques. A single band of protein having a molecular weight of 30,000 was found using dodecyl sulfate-polyacrylamide gel electrophoresis. Antisera were generated in rabbits against the purified enzyme and demonstrated to be monospecific by Ouchterlony immunodiffusion against crude homogenates from hydrocortisone-induced and uninduced C6 cells. All of the radioactivity in immunoprecipitates from (/sup 3/H)leucine-labeled cells co-migrated with purified glycerol phosphate dehydrogenase. The amount of radioactivity precipitated was directly proportional to the amount of labeled glycerol phosphate dehydrogenase present, indicating that the assay could be used to quantitate newly synthesized glycerol phosphate dehydrogenase molecules. Using these techniques, the induction of glycerol phosphate dehydrogenase activity by hydrocortisone in the C6 glioma cell line was shown to be due to an increase in the rate of synthesis of the enzyme. Analysis of the kinetics of induction and deinduction supports the above conclusion and suggests that there is essentially no change in the rate of degradation of glycerol phosphate dehydrogenase in the presence and absence of hormone.

• Research Organization: Univ. of California, Los Angeles
• OSTI ID: 6803755
• Journal Information: J. Biol. Chem.; (United States), Journal Name: J. Biol. Chem.; (United States) Vol. 253:23; ISSN JBCHA
• Country of Publication: United States
• Language: English

Related Subjects

550200* -- Biochemistry
59 BASIC BIOLOGICAL SCIENCES
ADRENAL HORMONES
ANIMAL CELLS
ANIMALS
BETA DECAY RADIOISOTOPES
BETA-MINUS DECAY RADIOISOTOPES
BIOCHEMICAL REACTION KINETICS
BIOSYNTHESIS
BODY
BRAIN
CELL CULTURES
CENTRAL NERVOUS SYSTEM
CORTICOSTEROIDS
DEHYDROGENASES
ENZYMES
GLUCOCORTICOIDS
HOMEOSTASIS
HORMONES
HYDROCORTISONE
HYDROGEN ISOTOPES
HYDROXY COMPOUNDS
ISOTOPE APPLICATIONS
ISOTOPES
KETONES
KINETICS
LIGHT NUCLEI
MAMMALS
MUSCLES
NERVOUS SYSTEM
NUCLEI
ODD-EVEN NUCLEI
ORGANIC COMPOUNDS
ORGANS
OXIDOREDUCTASES
PREGNANES
RABBITS
RADIOISOTOPES
RATS
REACTION KINETICS
RODENTS
STEROID HORMONES
STEROIDS
SYNTHESIS
TRACER TECHNIQUES
TRITIUM
TUMOR CELLS
VERTEBRATES
YEARS LIVING RADIOISOTOPES