Inhibitors of Protein Geranylgeranyltransferase I and Rab Geranylgeranyltransferase Identified from a Library of Allenoate-derived Compounds

Watanabe, Masaru; Fiji, Hannah D.G.; Guo, Lea; Chan, Lai N.; Kinderman, Sape S.; Slamon, Dennis J.; Kwon, Ohyun; Tamanoi, Fuyuhiko

doi:10.1074/jbc.m706229200

Cited by 81 publications

(78 citation statements)

References 42 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Furthermore, the present studies suggest that this behavior of NBD-FPP may be related to an inability to bind the putative second site on RGGT. Recently, another study reported on the identification of novel RGGT inhibitors (25). These inhibitors appear to act via a different mechanism, prob- ably binding to the peptide site as they exhibit competitive kinetics with respect to Rab and uncompetitive kinetics with respect to GGPP.…”

Section: Discussionmentioning

confidence: 99%

Phosphonocarboxylates Inhibit the Second Geranylgeranyl Addition by Rab Geranylgeranyl Transferase

Baron

Tavaré

Figueiredo

et al. 2009

Journal of Biological Chemistry

View full text Add to dashboard Cite

Rab geranylgeranyl transferase (RGGT) catalyzes the posttranslational geranylgeranyl (GG) modification of (usually) two C-terminal cysteines in Rab GTPases. Here we studied the mechanism of the Rab geranylgeranylation reaction by bisphosphonate analogs in which one phosphonate group is replaced by a carboxylate (phosphonocarboxylate, PC). The phosphonocarboxylates used were 3-PEHPC, which was previously reported, and 2-hydroxy-3-imidazo[1,2-a]pyridin-3-yl-2-phosphonopropionic acid ((؉)-3-IPEHPC), a >25-fold more potent related compound as measured by both IC 50 and K i . (؉)-3-IPEHPC behaves as a mixed-type inhibitor with respect to GG pyrophosphate (GGPP) and an uncompetitive inhibitor with respect to Rab substrates. We propose that phosphonocarboxylates prevent only the second GG transfer onto Rabs based on the following evidence. First, geranylgeranylation of Rab proteins ending with a single cysteine motif such as CAAX, is not affected by the inhibitors, either in vitro or in vivo. Second, the addition of an -AAX sequence onto Rab-CC proteins protects the substrate from inhibition by the inhibitors. Third, we demonstrate directly that in the presence of (؉)-3-IPEHPC, Rab-CC and Rab-CXC proteins are modified by only a single GG addition. The presence of (؉)-3-IPEHPC resulted in a preference for the Rab N-terminal cysteine to be modified first, suggesting an order of cysteine geranylgeranylation in RGGT catalysis. Our results further suggest that the inhibitor binds to a site distinct from the GGPP-binding site on RGGT. We suggest that phosphonocarboxylate inhibitors bind to a GG-cysteine binding site adjacent to the active site, which is necessary to align the mono-GG-Rab for the second GG addition. These inhibitors may represent a novel therapeutic approach in Rab-mediated diseases.Most proteins of the Ras-like GTPase superfamily need to be post-translationally modified by prenyl groups to associate with cellular membranes and to activate downstream effectors (1). Protein prenylation involves the formation of a thioether link between conserved C-terminal cysteines in protein substrates and farnesyl pyrophosphate (FPP) 7 or geranylgeranyl pyrophosphate (GGPP) (2, 3). These prenyl pyrophosphates, which originate from the mevalonate pathway are utilized by three different prenyltransferase enzymes (2, 3). Farnesyl transferase (FT) and geranylgeranyl transferase type I (GGT-I) transfer FPP or GGPP, respectively, to a cysteine residue in the context of a C-terminal CAAX motif, where C is a cysteine, A is an aliphatic residue, and X is any amino acid. X contributes significantly to substrate specificity in FT and GGT-I (2). Rab geranylgeranyl transferase (RGGT) is distinct from FT and GGT-I in that it specifically recognizes Rab proteins, which vary in their C terminus containing CCXX, XCXC, XXCC, CCXXX, CAAX, and other motifs (3). RGGT exhibits exquisite specificity for Rabs due to the strict requirement of Rab escort protein (REP), which is a general Rab-GDP binding protein (4). RGGT is a heterodimeric enzyme ...

show abstract

Section: Discussionmentioning

confidence: 99%

Phosphonocarboxylates Inhibit the Second Geranylgeranyl Addition by Rab Geranylgeranyl Transferase

Baron

Tavaré

Figueiredo

et al. 2009

Journal of Biological Chemistry

View full text Add to dashboard Cite

show abstract

“…Cell viability was determined with Cell Counting Kit-8 -(Dojindo) as described previously (40). Briefly, cells (2.5 × 10 3 for CGL4 and CGL4/gt3) were plated onto 96-well plates and treated with the appropriate inhibitor as indicated in figure legends.…”

Section: Cell Viability Analysismentioning

confidence: 99%

DNA Damage–Induced Modulation of GLUT3 Expression Is Mediated through p53-Independent Extracellular Signal-Regulated Kinase Signaling in HeLa Cells

Watanabe

Naraba

Sakyo

et al. 2010

Molecular Cancer Research

Self Cite

View full text Add to dashboard Cite

Many cancer cells exhibit increased rates of uptake and metabolism of glucose compared with normal cells. Glucose uptake in mammalian cells is mediated by the glucose transporter (GLUT) family. Here, we report that DNA-damaging anticancer agents such as Adriamycin and etoposide suppressed the expression of GLUT3, but not GLUT1, in HeLa cells and a tumorigenic HeLa cell hybrid. Suppression of GLUT3 expression determined by the real-time PCR was also evident with another DNA-damaging agent, camptothecin, which reduced the promoter's activity as determined with a luciferase-linked assay. The suppression by these agents seemed to be induced independently of p53, and it was evident when wild-type p53 was overproduced in these cells. In contrast, the mitogen-activated protein kinase/extracellular signal regulated kinase (MAPK/ERK) kinase (MEK) inhibitor U0126 (but not the phosphoinositide 3-kinase inhibitor LY294002) prevented the drug-induced suppression as determined by reverse transcription-PCR and promoter assays. Furthermore, overexpression of GLUT3 in HeLa cell hybrids increased resistance to these drugs, whereas depletion of the gene by small interfering RNA rendered the cells more sensitive to the drugs, decreasing glucose consumption. The results suggest that DNA-damaging agents reduce GLUT3 expression in cancer cells through activation of the MEK-ERK pathway independently of p53, leading to cell death or apoptosis. The findings may contribute to the development of new chemotherapeutic drugs based on the GLUT3-dependent metabolism of glucose.

show abstract

“…That finding prompted both pharmaceutical companies and academic laboratories to develop GGTase-I inhibitors (GGTIs) (26), which have shown promise in preclinical studies (27)(28)(29)(30)(31). The rationale for inhibiting GGTase-I is supported by genetic studies in mice: Inactivating the gene for the β-subunit of GGTase-I (Pggt1b) reduced tumor formation and prolonged survival in mice with K-RAS-induced lung cancer (32).…”

mentioning

confidence: 99%

Targeting the protein prenyltransferases efficiently reduces tumor development in mice with K-RAS-induced lung cancer

Liu

Sjögren

Karlsson

et al. 2010

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

View full text Add to dashboard Cite

RAS and RHO proteins, which contribute to tumorigenesis and metastasis, undergo posttranslational modification with an isoprenyl lipid by protein farnesyltransferase (FTase) or protein geranylgeranyltransferase-I (GGTase-I). Inhibitors of FTase and GGTase-I were developed to block RAS-induced malignancies, but their utility has been difficult to evaluate because of off-target effects, drug resistance, and toxicity. Moreover, the impact of FTase deficiency and combined FTase/ GGTase-I deficiency has not been evaluated with genetic approaches. We found that inactivation of FTase eliminated farnesylation of HDJ2 and H-RAS, prevented H-RAS targeting to the plasma membrane, and blocked proliferation of primary and K-RAS G12D -expressing fibroblasts. FTase inactivation in mice with K-RAS-induced lung cancer reduced tumor growth and improved survival, similar to results obtained previously with inactivation of GGTase-I. Simultaneous inactivation of FTase and GGTase-I markedly reduced lung tumors and improved survival without apparent pulmonary toxicity. These data shed light on the biochemical and therapeutic importance of FTase and suggest that simultaneous inhibition of FTase and GGTase-I could be useful in cancer therapeutics.mouse models | non-small-cell lung cancer | protein farnesyltransferase | protein geranylgeranyltransferase type I

show abstract

Inhibitors of Protein Geranylgeranyltransferase I and Rab Geranylgeranyltransferase Identified from a Library of Allenoate-derived Compounds

Cited by 81 publications

References 42 publications

Phosphonocarboxylates Inhibit the Second Geranylgeranyl Addition by Rab Geranylgeranyl Transferase

Phosphonocarboxylates Inhibit the Second Geranylgeranyl Addition by Rab Geranylgeranyl Transferase

DNA Damage–Induced Modulation of GLUT3 Expression Is Mediated through p53-Independent Extracellular Signal-Regulated Kinase Signaling in HeLa Cells

Targeting the protein prenyltransferases efficiently reduces tumor development in mice with K-RAS-induced lung cancer

Contact Info

Product

Resources

About