Determination of /sup 35/S-aminoacyl-transfer ribonucleic acid specific radioactivity in small tissue samples
Rate determination of protein synthesis utilizing tracer amino acid incorporation requires accurate assessment of the specific radioactivity of the labeled precursor aminoacyl-tRNA pool. Previously published methods presumably useful for the measurement of any aminoacyl-tRNA were unsuccessful when applied to (/sup 35/S)methionine, due to the unique chemical properties of this amino acid. Herein we describe modifications of these methods necessary for the measurement of /sup 35/S-aminoacyl-tRNA specific radioactivity from small tissue samples incubated in the presence of (/sup 35/S)methionine. The use of (/sup 35/S)methionine of high specific radioactivity enables analysis of the methionyl-tRNA from less than 100 mg of tissue. Conditions for optimal recovery of /sup 35/S-labeled dansyl-amino acid derivatives are presented and possible applications of this method are discussed.
- Research Organization:
- Northwestern Univ. School of Medicine, Chicago, IL
- OSTI ID:
- 5801919
- Journal Information:
- Anal. Biochem.; (United States), Vol. 118:1
- Country of Publication:
- United States
- Language:
- English
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Related Subjects
METHIONINE
MEASURING METHODS
RADIOASSAY
PROTEINS
BIOSYNTHESIS
SULFUR COMPOUNDS
DOUBLE LABELLING
TRANSFER RNA
TRITIUM COMPOUNDS
BIOCHEMICAL REACTION KINETICS
EXPERIMENTAL DATA
IN VITRO
ISOMERIC NUCLEI
MYOCARDIUM
RABBITS
SULFUR 35
TRACER TECHNIQUES
UPTAKE
AMINO ACIDS
ANIMALS
BETA DECAY RADIOISOTOPES
BETA-MINUS DECAY RADIOISOTOPES
BODY
CARBOXYLIC ACIDS
CARDIOVASCULAR SYSTEM
DATA
DAYS LIVING RADIOISOTOPES
DRUGS
EVEN-ODD NUCLEI
HEART
INFORMATION
ISOTOPE APPLICATIONS
ISOTOPES
KINETICS
LABELLED COMPOUNDS
LABELLING
LIGHT NUCLEI
LIPOTROPIC FACTORS
MAMMALS
MUSCLES
NUCLEI
NUCLEIC ACIDS
NUMERICAL DATA
ORGANIC ACIDS
ORGANIC COMPOUNDS
ORGANIC SULFUR COMPOUNDS
ORGANS
RADIOISOTOPES
REACTION KINETICS
RNA
SULFUR ISOTOPES
SYNTHESIS
VERTEBRATES
550201* - Biochemistry- Tracer Techniques