Inhibition of purified p21ras farnesyl: protein transferase by Cys-AAX tetrapeptides

Yuval Reiss, Joseph L. Goldstein, Miguel C. Seabra, Patrick J. Casey, Michael S. Brown

Research output: Contribution to journalArticle

674 Citations (Scopus)

Abstract

We report the identification, purification, and characterization of a farnesyl:protein transferase that transfers the farnesyl moiety from farnesyl pyrophosphate to a cysteine in p21ras proteins. The enzyme was purified ∼60,000-fold from rat brain cytosol through use of a chromatography step based on the enzyme's ability to bind to a hexapeptide containing the consensus sequence (Cys-AAX) for farnesylation. The purified enzyme migrated on gel filtration chromatography with an apparent molecular weight of 70,000-100,000. High resolution SDS-polyacrylamide gels showed two closely spaced ∼50 kd protein bands in the final preparation. The enzyme was inhibited competitively by peptides as short as 4 residues that contained the Cys-AAX motif. These peptides acted as alternative substrates that competed with p21H-ras for farnesylation. Effective peptides included the COOH-terminal sequences of all known p21ras proteins as well as those of lamin A and B.

Original languageEnglish (US)
Pages (from-to)81-88
Number of pages8
JournalCell
Volume62
Issue number1
DOIs
StatePublished - Jul 13 1990

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Transferases
Prenylation
Enzymes
Chromatography
Peptides
Proteins
Lamin Type B
Lamin Type A
Consensus Sequence
Cytosol
Purification
Gel Chromatography
Cysteine
Rats
Brain
Molecular Weight
Gels
Molecular weight
Substrates

ASJC Scopus subject areas

  • Cell Biology
  • Molecular Biology

Cite this

Inhibition of purified p21ras farnesyl : protein transferase by Cys-AAX tetrapeptides. / Reiss, Yuval; Goldstein, Joseph L.; Seabra, Miguel C.; Casey, Patrick J.; Brown, Michael S.

In: Cell, Vol. 62, No. 1, 13.07.1990, p. 81-88.

Research output: Contribution to journalArticle

Reiss, Yuval ; Goldstein, Joseph L. ; Seabra, Miguel C. ; Casey, Patrick J. ; Brown, Michael S. / Inhibition of purified p21ras farnesyl : protein transferase by Cys-AAX tetrapeptides. In: Cell. 1990 ; Vol. 62, No. 1. pp. 81-88.
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