Prenylated Proteins: Synthesis of Geranylgeranylcysteine and Identification of this Thioether Amino Acid as a Component of Proteins in CHO Cells

Prenylated proteins, labeled in the isoprenoid residue by growing CHO cells in medium containing [5-3H]mevalonate, were degraded by three different proteolytic procedures, enzymatic or alkaline hydrolysis as well as hydrazinolysis. The products thus obtained were analyzed by HPLC with chemically pre...

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Bibliographic Details
Published in:Proceedings of the National Academy of Sciences - PNAS 1990-10, Vol.87 (19), p.7352-7354
Main Authors: Epstein, W. W., Lever, D. C., Rilling, H. C.
Format: Article
Language:English
Subjects:
Amino acids
Animals
Biological and medical sciences
Carbohydrates
Cell Line
CHO cells
Chromatography, High Pressure Liquid
Cricetinae
Cricetulus
Cysteine - analogs & derivatives
Cysteine - analysis
Cysteine - chemical synthesis
Cysteine - metabolism
Diterpenes - analysis
Diterpenes - chemical synthesis
Diterpenes - metabolism
Elution
Female
Fundamental and applied biological sciences. Psychology
geranylgeranylcysteine
Hydrazines
Hydrolysis
Molecular and cellular biology
Molecular genetics
Ovary
Peptide Fragments - isolation & purification
Product labeling
Proteins
Proteins - isolation & purification
Proteins - metabolism
Sulfides
Terpenoids
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Summary:Prenylated proteins, labeled in the isoprenoid residue by growing CHO cells in medium containing [5-3H]mevalonate, were degraded by three different proteolytic procedures, enzymatic or alkaline hydrolysis as well as hydrazinolysis. The products thus obtained were analyzed by HPLC with chemically prepared all-trans-geranylgeranylcysteine as a standard. About 10% of the radioactive products released by each lytic procedure showed the same chromatographic properties as geranylgeranylcysteine. This verifies the earlier conclusion, based on less-direct evidence, that this thioether derivative of cysteine is a component of naturally occurring proteins. The finding of this modified amino acid as a product of hydrazinolysis indicates that it is a carboxyl-terminal amino acid and that it is not carboxyl-methylated.
ISSN:0027-8424
1091-6490
DOI:10.1073/pnas.87.19.7352