Initiation and elongation of protein synthesis: their relative rates and sensitivities to inhibition in Chinese hamster ovary cells determined by a modified Edman procedure
The relative rates of initiation and elongation of protein synthesis are determined in exponentially growing cells from the incorporation of (/sup 35/S)methionine into N-terminal and internal positions of growing peptide chains by a modified Edman degradation. Sequential samples of labeled cells are precipitated into filter paper supports with trichloroacetic acid. Large groups of samples can then be incubated in bulk with phenylisothiocyanate. Individual samples are treated with trifluoroacetic acid, and the derived N-terminal amino acids extracted from the filter paper with ethylene dichloride. The percentage of incorporation which is N-terminal varies with the purity of the (/sup 35/S)methionine. Elevated temperature and hypertonicity, inhibitors of initiation, preferentially block incorporation into the N-terminal fraction (initiation) but allow continued incorporation into internal positions of previously initiated peptides. Puromycin inhibited incorporation into both fractions, as expected for an inhibitor of elongation.
- Research Organization:
- Case Western Reserve Univ., Cleveland
- OSTI ID:
- 7347588
- Journal Information:
- Anal. Biochem.; (United States), Vol. 73:1
- Country of Publication:
- United States
- Language:
- English
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Related Subjects
ANIMAL CELLS
BIOCHEMICAL REACTION KINETICS
BIOSYNTHESIS
INHIBITION
PROTEINS
HAMSTERS
METHIONINE
OVARIES
PEPTIDES
SULFUR 35
AMINO ACIDS
ANIMALS
BETA DECAY RADIOISOTOPES
BETA-MINUS DECAY RADIOISOTOPES
BODY
CARBOXYLIC ACIDS
DAYS LIVING RADIOISOTOPES
EVEN-ODD NUCLEI
FEMALE GENITALS
GONADS
ISOTOPES
KINETICS
LIGHT NUCLEI
LIPOTROPIC FACTORS
MAMMALS
NUCLEI
ORGANIC ACIDS
ORGANIC COMPOUNDS
ORGANIC SULFUR COMPOUNDS
ORGANS
RADIOISOTOPES
REACTION KINETICS
RODENTS
SULFUR ISOTOPES
SYNTHESIS
VERTEBRATES
550200* - Biochemistry