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Characterization of the increased synthesis of rat renal glutaminase during metabolic acidosis

Thesis/Dissertation ·
OSTI ID:7140789

The relative rates of glutaminase synthesis were determined by comparing the amount of (/sup 35/S)methionine incorporated into specific immunoprecipitates with that incorporated into total protein. In a normal animal, the rate of glutaminase synthesis constitutes 0.04% of the total protein synthesis. During onset of acidosis, the relative rate of synthesis increases more rapidly than the appearance of increased glutaminase activity. Recovery from chronic acidosis results in a rapid decrease in the relative rate of glutaminase synthesis, but a gradual decrease in glutaminase activity. From the decrease in activity that occurs upon recovery from acidosis, the tube half-life for the glutaminase was estimated to be 3 days. The reticulocyte lysate was used to compare the level of translatable glutaminase mRNA that is present in rat kidney following translatable glutaminase mRNA that is present in rat kidney following onset of acidosis. The initial translate of glutaminase was previously identified as a 72,000 dalton protein that is 4000 daltons larger than the mitochondrial glutaminase. The amount of (/sup 35/S)methionine incorporated into the glutaminase was determined by densitometric tracing of specific immunoprecipitates. The relative rate of glutaminase synthesis was determined by comparing this value with the amount of (/sup 35/S) methionine incorporated into total protein. In order to obtain a more specific immunoprecipitate of the in vitro translate, poly(A)/sup +/RNA was fractionated. The fractionated poly(A)/sup +/RNA was about 25-fold enriched in glutaminase mRNA. The size of glutaminase mRNA was estimated to be between 6.4 Kb and 6.8 Kb. The effect of alteration in acid-base balance on the level of mRNA coding for glutaminase was also determined by using fractionated poly(A)/sup +/RNA from the kidneys of normal, chronic acidotic and recovered rats for in vitro translation.

Research Organization:
Pittsburgh Univ., PA (USA)
OSTI ID:
7140789
Country of Publication:
United States
Language:
English

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Related Subjects

550201* -- Biochemistry-- Tracer Techniques
59 BASIC BIOLOGICAL SCIENCES
AMINE OXIDASES
AMINO ACIDS
ANIMALS
BETA DECAY RADIOISOTOPES
BETA-MINUS DECAY RADIOISOTOPES
BIOCHEMICAL REACTION KINETICS
BIOLOGICAL HALF-LIFE
BIOLOGICAL MATERIALS
BIOSYNTHESIS
BLOOD
BLOOD CELLS
BODY
BODY FLUIDS
CARBOXYLIC ACIDS
CELL CONSTITUENTS
DAYS LIVING RADIOISOTOPES
DISEASES
DRUGS
ENZYME ACTIVITY
ENZYMES
ERYTHROCYTES
EVEN-ODD NUCLEI
FRACTIONATION
GLUTAMIC ACID
ISOTOPE APPLICATIONS
ISOTOPES
KIDNEYS
KINETICS
LABELLED COMPOUNDS
LIGHT NUCLEI
LIPOTROPIC FACTORS
MAMMALS
MATERIALS
MESSENGER-RNA
METABOLIC DISEASES
METHIONINE
MITOCHONDRIA
MOLECULAR WEIGHT
NUCLEI
NUCLEIC ACIDS
ORGANIC ACIDS
ORGANIC COMPOUNDS
ORGANIC SULFUR COMPOUNDS
ORGANOIDS
ORGANS
OXIDOREDUCTASES
PROTEINS
RADIOISOTOPES
RATS
REACTION KINETICS
RETICULOCYTES
RNA
RODENTS
SEPARATION PROCESSES
SULFUR 35
SULFUR ISOTOPES
SYNTHESIS
TRACER TECHNIQUES
TRANSCRIPTION
VERTEBRATES