Sequence requirement for peptide recognition by rat brain p21ras protein farnesyltransferase

Yuval Reiss, Sarah J. Stradley, Lila M. Gierasch, Michael S. Brown, Joseph L. Goldstein

Research output: Contribution to journalArticle

302 Citations (Scopus)

Abstract

We tested 42 tetrapeptides for their ability to bind to the rat brain p21ras protein farnesyltransferase as estimated by their ability to compete with p21Ha-ras in a farnesyltransfer assay. Peptides with the highest affinity had the structure Cys-A1-A2-X, where positions A1 and A2 are occupied by aliphatic amino acids and position X is occupied by a COOH-terminal methionine, serine, or phenylalanine. Charged residues reduced affinity slightly at the A1 position and much more drastically at the A2 and X positions. Effective inhibitors included tetrapeptides corresponding to the COOH termini of all animal cell proteins known to be farnesylated. In contrast, the tetrapeptide Cys-Ala-Ile-Leu (CAIL), which corresponds to the COOH termini of several neural guanine nucleotide binding (G) protein γ subunits, did not compete in the farnesyl-transfer assay. Inasmuch as several of these proteins are geranylgeranylated, the data suggest that the two isoprenes (farnesyl and geranylgeranyl) are transferred by different enzymes. A biotinylated heptapeptide corresponding to the COOH terminus of p21Ki-rasB was farnesylated, suggesting that at least some of the peptides serve as substrates for the transferase. The data are consistent with a model in which a hydrophobic pocket in the protein farnesyltransferase recognizes tetrapeptides through interactions with the cysteine and the last two amino acids.

Original languageEnglish (US)
Pages (from-to)732-736
Number of pages5
JournalProceedings of the National Academy of Sciences of the United States of America
Volume88
Issue number3
StatePublished - 1991

Fingerprint

varespladib methyl
Peptides
Brain
Amino Acids
Guanine Nucleotides
Protein Subunits
Transferases
Phenylalanine
GTP-Binding Proteins
Methionine
Serine
Cysteine
Carrier Proteins
Proteins
Fatty Acids
Enzymes
p21(ras) farnesyl-protein transferase

Keywords

  • Covalent modification
  • Enzyme inhibition
  • Mevalonate
  • Prenylation
  • Tetrapeptides

ASJC Scopus subject areas

  • General
  • Genetics

Cite this

Sequence requirement for peptide recognition by rat brain p21ras protein farnesyltransferase. / Reiss, Yuval; Stradley, Sarah J.; Gierasch, Lila M.; Brown, Michael S.; Goldstein, Joseph L.

In: Proceedings of the National Academy of Sciences of the United States of America, Vol. 88, No. 3, 1991, p. 732-736.

Research output: Contribution to journalArticle

@article{274a86a8cc494b00a6e41bbf3747d2ff,
title = "Sequence requirement for peptide recognition by rat brain p21ras protein farnesyltransferase",
abstract = "We tested 42 tetrapeptides for their ability to bind to the rat brain p21ras protein farnesyltransferase as estimated by their ability to compete with p21Ha-ras in a farnesyltransfer assay. Peptides with the highest affinity had the structure Cys-A1-A2-X, where positions A1 and A2 are occupied by aliphatic amino acids and position X is occupied by a COOH-terminal methionine, serine, or phenylalanine. Charged residues reduced affinity slightly at the A1 position and much more drastically at the A2 and X positions. Effective inhibitors included tetrapeptides corresponding to the COOH termini of all animal cell proteins known to be farnesylated. In contrast, the tetrapeptide Cys-Ala-Ile-Leu (CAIL), which corresponds to the COOH termini of several neural guanine nucleotide binding (G) protein γ subunits, did not compete in the farnesyl-transfer assay. Inasmuch as several of these proteins are geranylgeranylated, the data suggest that the two isoprenes (farnesyl and geranylgeranyl) are transferred by different enzymes. A biotinylated heptapeptide corresponding to the COOH terminus of p21Ki-rasB was farnesylated, suggesting that at least some of the peptides serve as substrates for the transferase. The data are consistent with a model in which a hydrophobic pocket in the protein farnesyltransferase recognizes tetrapeptides through interactions with the cysteine and the last two amino acids.",
keywords = "Covalent modification, Enzyme inhibition, Mevalonate, Prenylation, Tetrapeptides",
author = "Yuval Reiss and Stradley, {Sarah J.} and Gierasch, {Lila M.} and Brown, {Michael S.} and Goldstein, {Joseph L.}",
year = "1991",
language = "English (US)",
volume = "88",
pages = "732--736",
journal = "Proceedings of the National Academy of Sciences of the United States of America",
issn = "0027-8424",
number = "3",

}

TY - JOUR

T1 - Sequence requirement for peptide recognition by rat brain p21ras protein farnesyltransferase

AU - Reiss, Yuval

AU - Stradley, Sarah J.

AU - Gierasch, Lila M.

AU - Brown, Michael S.

AU - Goldstein, Joseph L.

PY - 1991

Y1 - 1991

N2 - We tested 42 tetrapeptides for their ability to bind to the rat brain p21ras protein farnesyltransferase as estimated by their ability to compete with p21Ha-ras in a farnesyltransfer assay. Peptides with the highest affinity had the structure Cys-A1-A2-X, where positions A1 and A2 are occupied by aliphatic amino acids and position X is occupied by a COOH-terminal methionine, serine, or phenylalanine. Charged residues reduced affinity slightly at the A1 position and much more drastically at the A2 and X positions. Effective inhibitors included tetrapeptides corresponding to the COOH termini of all animal cell proteins known to be farnesylated. In contrast, the tetrapeptide Cys-Ala-Ile-Leu (CAIL), which corresponds to the COOH termini of several neural guanine nucleotide binding (G) protein γ subunits, did not compete in the farnesyl-transfer assay. Inasmuch as several of these proteins are geranylgeranylated, the data suggest that the two isoprenes (farnesyl and geranylgeranyl) are transferred by different enzymes. A biotinylated heptapeptide corresponding to the COOH terminus of p21Ki-rasB was farnesylated, suggesting that at least some of the peptides serve as substrates for the transferase. The data are consistent with a model in which a hydrophobic pocket in the protein farnesyltransferase recognizes tetrapeptides through interactions with the cysteine and the last two amino acids.

AB - We tested 42 tetrapeptides for their ability to bind to the rat brain p21ras protein farnesyltransferase as estimated by their ability to compete with p21Ha-ras in a farnesyltransfer assay. Peptides with the highest affinity had the structure Cys-A1-A2-X, where positions A1 and A2 are occupied by aliphatic amino acids and position X is occupied by a COOH-terminal methionine, serine, or phenylalanine. Charged residues reduced affinity slightly at the A1 position and much more drastically at the A2 and X positions. Effective inhibitors included tetrapeptides corresponding to the COOH termini of all animal cell proteins known to be farnesylated. In contrast, the tetrapeptide Cys-Ala-Ile-Leu (CAIL), which corresponds to the COOH termini of several neural guanine nucleotide binding (G) protein γ subunits, did not compete in the farnesyl-transfer assay. Inasmuch as several of these proteins are geranylgeranylated, the data suggest that the two isoprenes (farnesyl and geranylgeranyl) are transferred by different enzymes. A biotinylated heptapeptide corresponding to the COOH terminus of p21Ki-rasB was farnesylated, suggesting that at least some of the peptides serve as substrates for the transferase. The data are consistent with a model in which a hydrophobic pocket in the protein farnesyltransferase recognizes tetrapeptides through interactions with the cysteine and the last two amino acids.

KW - Covalent modification

KW - Enzyme inhibition

KW - Mevalonate

KW - Prenylation

KW - Tetrapeptides

UR - http://www.scopus.com/inward/record.url?scp=0026061739&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0026061739&partnerID=8YFLogxK

M3 - Article

C2 - 1992464

AN - SCOPUS:0026061739

VL - 88

SP - 732

EP - 736

JO - Proceedings of the National Academy of Sciences of the United States of America

JF - Proceedings of the National Academy of Sciences of the United States of America

SN - 0027-8424

IS - 3

ER -