A protein geranylgeranyltransferase from bovine brain: implications for protein prenylation specificity.

Yokoyama, Kohei; Goodwin, Gary W.; Ghomashchi, Farideh; Glomset, John A.; Gelb, Michael H.

doi:10.1073/pnas.88.12.5302

Cited by 224 publications

(204 citation statements)

References 35 publications

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“…As shown in Figure 9( [2,18]; (ii) DPR1 protein purified from E. coli can reconstitute FTase when added to the extracts prepared from dprl mutant cells [18]; (iii) sequences of DPR1 and RAM2 proteins share a low, but significant, similarity with ,-and a-subunits respectively of the mammalian FTase [19,20]; (iv) extracts of E. coli cells expressing both DPRI and RAM2 exhibit FTase activity [21]. What has been lacking in these studies is the direct demonstration of [10] reported similar findings using the mammalian FTase and peptide substrates. Thus changing the C-terminal amino acid of an FTase substrate to leucine alters the substrate to be preferentially utilized by GGTase, but does not exclude it being used as an FTase substrate.…”

Section: Peptide Inhibitionmentioning

confidence: 99%

“…identity between DPR1 and the f-subunit of the mammalian Another type of protein prenylation involves the addition of a FTase as well as between RAM2 and the a-subunit of the geranylgeranyl group, and protein geranylgeranyltransferases mammalian FTase [19,20]. In addition, FTase activity was (GGTases) are responsible for this type of modification [6][7][8][9][10][11].…”

Section: Introductionmentioning

confidence: 99%

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Purified yeast protein farnesyltransferase is structurally and functionally similar to its mammalian counterpart

Gomez

Goodman

Tripathy

et al. 1993

Biochemical Journal

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Protein farnesyltransferase (FTase) catalyses the addition of a farnesyl group to a cysteine within the so-called ‘CAAX box’ at the C-terminus of various proteins. In the present paper we report purification of Saccharomyces cerevisiae FTase to near-homogeneity. This was accomplished by constructing a yeast strain overproducing FTase approx. 100-fold. The purified enzyme was a heterodimer of approx. 90 kDa and consisted of 43 kDa and 34 kDa subunits. The 43 kDa subunit was shown to be the product of the DPR1 gene by using antibody raised against baculovirus-produced DPR1 polypeptide. The purified enzyme required Mg2+, showed a pH optimum of 7.8 and was most active at 50 degrees C. The Km values for farnesyl pyrophosphate and GST-CIIS (glutathione S-transferase fused to the C-terminal 12 amino acids of yeast RAS2 protein), KmFpp and KmGST CIIS, were 8.1 and 5.1 microM respectively. The enzyme was capable of farnesylating GST-CIIL (the same as GST-CIIS, except that the C-terminal serine is changed to leucine), a substrate protein for the enzyme geranylgeranyltransferase, although with a higher apparent Km than for GST-CIIS. Like its mammalian counterpart, yeast FTase activity was inhibited by peptides containing the C-terminal CAAX sequence (that is, one where C = cysteine, A = aliphatic amino acid and X = any amino acid). These results provide direct evidence for the idea that the yeast and mammalian FTases are structurally and functionally very similar.

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Section: Peptide Inhibitionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Purified yeast protein farnesyltransferase is structurally and functionally similar to its mammalian counterpart

Gomez

Goodman

Tripathy

et al. 1993

Biochemical Journal

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“…Genetic and biochemical studies on protein prenylation have revealed three distinct enzymes, each consisting of evolutionarily conserved subunits (Chen et al, 1991b;Kohl et al, 1991). Geranylgeranyl transferase I (GGTase I or CAAX GGTase) Seabra et al, 1991;Yokoyama et al, 1991;Moomaw and Casey, 1992) and farnesyl transferase (FTase) (Reiss et al, 1990(Reiss et al, , 1991aChen et al, 1991a;Seabra et al, 1991) modify proteins containing the Cys-Ali-Ali-X sequence Yokoyama et al, 1991;Reiss et al, 1991b;Kinsella et al, 1991), form heterodimers, and share a common a subunit (Seabra et al, 1991). The third prenyltransferase, named GGTase II or Rab GGTase (Seabra et al, 1992), modifies proteins that terminate with GGCC or CXC and consists of three nonidentical subunits (Seabra et al, 1992).…”

Section: Introductionmentioning

confidence: 99%

“…In vitro study of substrate specificity using partially purified FTase and GGTase I is basically consistent with this rule. However, the FTase can modify proteins with C-terminal leucine to a limited extent when a high concentration of the protein substrate is used (Yokoyama et al, 1991). This in vitro cross-specificity may be explained by the fact that the two enzymes not only share a common a subunit but have :3 subunits that, though not identical, are nevertheless similar to each other.…”

Section: Introductionmentioning

confidence: 99%

Suppression of yeast geranylgeranyl transferase I defect by alternative prenylation of two target GTPases, Rho1p and Cdc42p.

Ohya

Qadota

Anraku

et al. 1993

MBoC

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Geranylgeranyl transferase I (GGTase I), which modifies proteins containing the sequence Cys-Ali-Ali-Leu (Ali: aliphatic) at their C-termini, is indispensable for growth in the budding yeast Saccharomyces cerevisiae. We report here that GGTase I is no longer essential when Rho1p and Cdc42p are simultaneously overproduced. The lethality of a GGTase I deletion is most efficiently suppressed by provision of both Rho1p and Cdc42p with altered C-terminal sequences (Cys-Ali-Ali-Met) corresponding to the C-termini of substrates of farnesyl transferase (FTase). Under these circumstances, the FTase, normally not essential for growth of yeast, becomes essential.

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“…These modifications include derivitization of the cysteine sulfhydryl with an isoprenoid moiety followed by the endoproteolytic removal of the -AAX tripeptide and methylation of the cysteine ␣-carboxyl group. When the X amino acid is S, C, Q, or M, a 15-carbon farnesyl residue is attached in thioether linkage to the cysteine (3), whereas when X is a leucine, a 20-carbon geranylgeranyl residue is found instead (4).…”

mentioning

confidence: 99%

In Vitro Assay and Characterization of the Farnesylation-dependent Prelamin A Endoprotease

Kilic

Dalton

Burrell

et al. 1997

Journal of Biological Chemistry

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The 72-kDa nuclear lamina protein lamin A is synthesized as a 74-kDa farnesylated precursor. Conversion of this precursor to mature lamin A appears to be mediated by a specific endoprotease. Prior studies of overexpressed wild-type and mutant lamin A proteins in cultured cells have indicated that the precursor possesses the typical carboxyl-terminal S-farnesylated, cysteine methyl ester and that farnesylation is required for endoproteolysis to occur. In this report, we describe the synthesis of an S-farnesyl, cysteinyl methyl ester peptide corresponding to the carboxyl-terminal 18 amino acid residues of human prelamin A. This peptide acts as a substrate for the prelamin A endoprotease in vitro, with cleavage of the synthetic peptide at the expected site between Tyr 657 and Leu 658 . Endoproteolytic cleavage requires the S-prenylated cysteine methyl ester and, in agreement with transfection studies, is more active with the farnesylated than geranylgeranylated cysteinyl substrate. N-Acetyl farnesyl methyl cysteine is shown to be a noncompetitive inhibitor of the enzyme. Taken together, these observations suggest that there is a specific farnesyl binding site on the enzyme which is not at the active site.Proteins with a CAAX consensus sequence at their carboxyl terminus undergo serial post-translational modifications of the cysteinyl residue (1, 2). These modifications include derivitization of the cysteine sulfhydryl with an isoprenoid moiety followed by the endoproteolytic removal of the -AAX tripeptide and methylation of the cysteine ␣-carboxyl group. When the X amino acid is S, C, Q, or M, a 15-carbon farnesyl residue is attached in thioether linkage to the cysteine (3), whereas when X is a leucine, a 20-carbon geranylgeranyl residue is found instead (4).The nuclear lamina is a thin, fibrous structure that lines the inner nuclear membrane and is believed to function in maintaining nuclear shape and volume (5) and may also be involved in the organization of chromatin in the interphase nucleus (6). In most mammalian cells, it consists of three class V intermediate filament proteins, lamins A, B, and C (5, 6). Prelamin A is the 74-kDa precursor of the 72-kDa nuclear lamin A protein (7).It possesses a CAAX box sequence (CSIM) (8, 9) and has been shown to be farnesylated in vitro (10) and in vivo (11). Despite the loss of the carboxyl-terminal 18 amino acids of prelamin A in its proteolytic conversion to lamin A, it nevertheless undergoes all of the reactions characteristic of other CAAX proteins (11). Experiments with mutants, in which the cysteine of the CAAX box is replaced by another amino acid, demonstrate that farnesylation is required for the maturation of prelamin A (12). These nonprenylated CAAX box mutants of prelamin A enter the nucleus, yet are not proteolytically processed and are not incorporated into the nuclear lamina. Similar results have been obtained with nonprenylated prelamin A produced by treating cultured mammalian cells with mevinolin (13,14) or inhibitors of protein farnesylation (15).Prela...

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A protein geranylgeranyltransferase from bovine brain: implications for protein prenylation specificity.

Cited by 224 publications

References 35 publications

Purified yeast protein farnesyltransferase is structurally and functionally similar to its mammalian counterpart

Purified yeast protein farnesyltransferase is structurally and functionally similar to its mammalian counterpart

Suppression of yeast geranylgeranyl transferase I defect by alternative prenylation of two target GTPases, Rho1p and Cdc42p.

In Vitro Assay and Characterization of the Farnesylation-dependent Prelamin A Endoprotease

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