Crystal Structure of Farnesyl Protein Transferase Complexed with a CaaX Peptide and Farnesyl Diphosphate Analogue

Strickland, Corey L.; Windsor, William; Syto, Rosalinda; Wang, Lynn; Bond, Richard; Wu, Zezhou; Schwartz, Jeffrey; Le, Hung V.; Beese, L.S.; Weber, Patricia C

doi:10.1021/bi981197z

Cited by 215 publications

(292 citation statements)

References 30 publications

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“…Sequence analysis revealed a deletion of 11 nucleotides in the third exon of the PFT͞PGGT-I␣ gene in plp-1. The deletion removes a conserved four amino acid motif (DAKH), which includes a putative substrate-binding site (40) and creates a stop codon predicted to result in a truncated protein of 146 amino acids (17.4 kDa). Sequencing of a PFT͞PGGT-I␣-specific RT-PCR product from plp-1 confirmed this deletion.…”

Section: Resultsmentioning

confidence: 99%

Enlarged meristems and delayed growth in plp mutants result from lack of CaaX prenyltransferases

Running

Lavy²,

Sternberg³

et al. 2004

Proc. Natl. Acad. Sci. U.S.A.

122

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Meristems require a myriad of intercellular signaling pathways for coordination of cell division within and between functional zones and clonal cell layers. This control of cell division ensures a constant availability of stem cells throughout the life span of the meristem while limiting overproliferation of meristematic cells and maintaining the meristem structure. We have undertaken a genetic screen to identify additional components of meristem signaling pathways. We identified pluripetala (plp) mutants based on their dramatically larger meristems and increased floral organ number. PLURIPETALA encodes the ␣-subunit shared between protein farnesyltransferase and protein geranylgeranyltransferase-I. plp mutants also have altered abscisic acid responses and overall much slower growth rate. plp is epistatic to mutations in the ␤-subunit of farnesyltransferase and shows a synergistic interaction with clavata3 mutants. plp mutants lead to insights into the mechanism of meristem homeostasis and provide a unique in vivo system for studying the functional role of prenylation in eukaryotes.

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Section: Resultsmentioning

confidence: 99%

Enlarged meristems and delayed growth in plp mutants result from lack of CaaX prenyltransferases

Running

Lavy²,

Sternberg³

et al. 2004

Proc. Natl. Acad. Sci. U.S.A.

122

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“…This observation is of particular interest in light of the fact that the negatively charged surface of FTase with which the polybasic domain interacts resides primarily within the ␣-subunit of the enzyme (38) that is common to both FTase and GGTase. Therefore, geranylgeranylated proteins that contain polybasic domains may be more resistant to Ras-competitive GGTIs than those that do not, suggesting that these proteins can also be usefully defined according to the presence or absence of a polybasic domain.…”

Section: Discussionmentioning

confidence: 99%

“…However, replacement of the polybasic domain with five consecutive glutamines (K-(Q 5 ); IC 50 ϭ 80 nM) while keeping the CVIM sequence intact reduces the Ras/ FTase association to levels approximating that of H-Ras (IC 50 ϭ 10 nM). Based on the crystal structure of FTase complexed with a K-Ras4B CAAX peptide and a farnesyl diphosphate analogue (38), it seems likely that increased affinity between FTase and Ras results from the ionic interaction of the Ras polybasic domain and an acidic region on the surface of FTase near the active site. Residual physical association between K-(Q 5 ) and FTase probably results from hydrogen-bonding interactions between the polar glutamine residues and this negatively charged region of FTase.…”

Section: Design and Construction Of Mutant Ras Proteins-in Order Tomentioning

confidence: 99%

High Affinity for Farnesyltransferase and Alternative Prenylation Contribute Individually to K-Ras4B Resistance to Farnesyltransferase Inhibitors

Fiordalisi

Johnson

Weinbaum

et al. 2003

Journal of Biological Chemistry

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Farnesyltransferase inhibitors (FTIs) block Ras farnesylation, subcellular localization and activity, and inhibit the growth of Ras-transformed cells. Although FTIs are ineffective against K-Ras4B, the Ras isoform most commonly mutated in human cancers, they can inhibit the growth of tumors containing oncogenic K-Ras4B, implicating other farnesylated proteins or suggesting distinct functions for farnesylated and for geranylgeranylated K-Ras, which is generated when farnesyltransferase is inhibited. In addition to bypassing FTI blockade through geranylgeranylation, K-Ras4B resistance to FTIs may also result from its higher affinity for farnesyltransferase. Using chimeric Ras proteins containing all combinations of Ras background, CAAX motif, and K-Ras polybasic domain, we show that either a polybasic domain or an alternatively prenylated CAAX renders Ras prenylation, Ras-induced Elk-1 activation, and anchorage-independent cell growth FTI-resistant. The polybasic domain alone increases the affinity of Ras for farnesyltransferase, implying independent roles for each K-Ras4B sequence element in FTI resistance. Using microarray analysis and colony formation assays, we confirm that K-Ras function is independent of the identity of the prenyl group and, therefore, that FTI inhibition of K-Ras transformed cells is likely to be independent of K-Ras inhibition. Our results imply that relevant FTI targets will lack both polybasic and potentially geranylgeranylated methionine-CAAX motifs.

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“…All three amino acid residues are located along one side of a hydrophobic pocket formed by the ␤-subunit (6). In particular, position 362 (position 361 in the rat enzyme) is believed to stabilize the peptide substrate binding (5,8).Inhibitors of farnesyltransferase (FTIs) have recently emerged as promising anti-cancer drugs (1, 2). FTIs block anchorage-independent growth of transformed cells and induce their morphological reversion (16 -20).…”

mentioning

confidence: 99%

mentioning

confidence: 99%

A Mutant Form of Human Protein Farnesyltransferase Exhibits Increased Resistance to Farnesyltransferase Inhibitors

Villar¹,

Urano²,

Guo³

et al. 1999

Journal of Biological Chemistry

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Protein farnesyltransferase (FTase) is a key enzyme responsible for the lipid modification of a large and important number of proteins including Ras. Recent demonstrations that inhibitors of this enzyme block the growth of a variety of human tumors point to the importance of this enzyme in human tumor formation. In this paper, we report that a mutant form of human FTase, Y361L, exhibits increased resistance to farnesyltransferase inhibitors, particularly a tricyclic compound, SCH56582, which is a competitive inhibitor of FTase with respect to the CAAX (where C is cysteine, A is an aliphatic amino acid, and X is the C-terminal residue that is preferentially serine, cysteine, methionine, glutamine or alanine) substrates. The Y361L mutant maintains FTase activity toward substrates ending with CIIS. However, the mutant also exhibits an increased affinity for peptides terminating with CIIL, a motif that is recognized by geranylgeranyltransferase I (GGTase I). The Y361L mutant also demonstrates activity with Ha-Ras and Cdc42Hs proteins, substrates of FTase and GGTase I, respectively. In addition, the Y361L mutant shows a marked sensitivity to a zinc chelator HPH-5 suggesting that the mutant has altered zinc coordination. These results demonstrate that a single amino acid change at a residue at the active site can lead to the generation of a mutant resistant to FTase inhibitors. Such a mutant may be valuable for the study of the effects of FTase inhibitors on tumor cells.Protein farnesyltransferase (FTase) 1 catalyzes the transfer of a farnesyl group onto a conserved cysteine residue four amino acids from the C terminus of a number of proteins, such as Ras, that are involved in cell growth and morphogenesis (1-4). This modification is critical for membrane association and subsequent protein-protein interactions of these proteins. FTase is a heterodimeric enzyme consisting of ␣-and ␤-subunits. The ␣-subunit of FTase is shared with the related prenyltransferase, protein geranylgeranyltransferase type I (GGTase I), whereas the ␤-subunits of FTase and GGTase I are approximately 30% homologous. The FTase enzyme recognizes the CAAX motif (C is cysteine, A is an aliphatic amino acid, and X is the C-terminal residue that is preferentially serine, cysteine, methionine, glutamine or alanine) that is found at the C termini of the substrate proteins. GGTase I enzyme also recognizes a CAAX motif; however, the terminal X amino acid is predominantly leucine or phenylalanine. This motif is referred to as the CAAL motif.The three-dimensional structures of the rat FTase enzyme have recently been resolved without substrate bound (5, 6), with bound substrate farnesyl pyrophosphate (FPP) (7), and with bound FPP and CAAX peptide analogs (8). In the structure without bound substrate, a non-cognate peptide provided by an adjacent ␤-subunit was modeled into the active site of the enzyme. The structure showed that the ␣-and ␤-subunits are largely composed of ␣-helices. The ␤-subunit forms a barrel-like structure, and one side of this barrel i...

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Crystal Structure of Farnesyl Protein Transferase Complexed with a CaaX Peptide and Farnesyl Diphosphate Analogue

Cited by 215 publications

References 30 publications

Enlarged meristems and delayed growth in plp mutants result from lack of CaaX prenyltransferases

Enlarged meristems and delayed growth in plp mutants result from lack of CaaX prenyltransferases

High Affinity for Farnesyltransferase and Alternative Prenylation Contribute Individually to K-Ras4B Resistance to Farnesyltransferase Inhibitors

A Mutant Form of Human Protein Farnesyltransferase Exhibits Increased Resistance to Farnesyltransferase Inhibitors

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