Photoaffinity labeling with GTP of viral p21 ras protein expressed in Escherichia coli
- PMID: 6323735
- PMCID: PMC255626
- DOI: 10.1128/JVI.50.2.343-351.1984
Photoaffinity labeling with GTP of viral p21 ras protein expressed in Escherichia coli
Abstract
The v-ras oncogene of Harvey murine sarcoma virus encodes a 21,000-dalton protein, p21, which mediates transformation produced by that virus. Previous work has shown that both p21v-rasH and the cellular homolog p21c-rasH appear to bind guanine nucleotides. We report here the expression in Escherichia coli of v-rasH to produce a biochemically active p21 fusion protein which retains both guanine nucleotide binding and autophosphorylating activity. Furthermore, direct interaction of this protein with GTP is unequivocally demonstrated by photoaffinity labeling it with [alpha-32P]GTP.
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