The Specific Binding of Small Molecule Isoprenoids to <i>rho</i>GDP Dissociation Inhibitor (<i>rho</i>GDI)

Mondal, Madhu Sudan; Wang, Zhaolin; Seeds, Andrew M.; Rando, Robert R.

doi:10.1021/bi991856n

Cited by 24 publications

(49 citation statements)

References 41 publications

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“…The weak binding observed in the absence of the methyl group is likely caused by the charge repulsion effect of the carboxylic acid group and the decreased hydrophobicity of farnesylated C185. Our results are in accordance with the previous findings showing that carboxymethylation of prenylated Rho peptide and catalytic subunits of PDE6 significantly increases their affinity for RhoGDI and PDEδ, respectively (33,36). Carboxymethylation is the only step in the posttranslational modification of the CaaX motif that is potentially reversible (37).…”

Section: Discussionsupporting

confidence: 93%

Structural basis of recognition of farnesylated and methylated KRAS4b by PDEδ

Dharmaiah

Bindu

Tran

et al. 2016

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

157

197

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Farnesylation and carboxymethylation of KRAS4b (Kirsten rat sarcoma isoform 4b) are essential for its interaction with the plasma membrane where KRAS-mediated signaling events occur. Phosphodiesterase-δ (PDEδ) binds to KRAS4b and plays an important role in targeting it to cellular membranes. We solved structures of human farnesylated–methylated KRAS4b in complex with PDEδ in two different crystal forms. In these structures, the interaction is driven by the C-terminal amino acids together with the farnesylated and methylated C185 of KRAS4b that binds tightly in the central hydrophobic pocket present in PDEδ. In crystal form II, we see the full-length structure of farnesylated–methylated KRAS4b, including the hypervariable region. Crystal form I reveals structural details of farnesylated–methylated KRAS4b binding to PDEδ, and crystal form II suggests the potential binding mode of geranylgeranylated–methylated KRAS4b to PDEδ. We identified a 5-aa-long sequence motif (Lys-Ser-Lys-Thr-Lys) in KRAS4b that may enable PDEδ to bind both forms of prenylated KRAS4b. Structure and sequence analysis of various prenylated proteins that have been previously tested for binding to PDEδ provides a rationale for why some prenylated proteins, such as KRAS4a, RalA, RalB, and Rac1, do not bind to PDEδ. Comparison of all four available structures of PDEδ complexed with various prenylated proteins/peptides shows the presence of additional interactions due to a larger protein–protein interaction interface in KRAS4b–PDEδ complex. This interface might be exploited for designing an inhibitor with minimal off-target effects.

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Section: Discussionsupporting

confidence: 93%

Structural basis of recognition of farnesylated and methylated KRAS4b by PDEδ

Dharmaiah

Bindu

Tran

et al. 2016

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

157

197

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“…3). Moreover, it was shown that the binding affinity of RhoGDI-1 for the farnesylated and geranylgeranylated moiety is not really important (42).…”

Section: Discussionmentioning

confidence: 99%

RhoA Prenylation Is Required for Promotion of Cell Growth and Transformation and Cytoskeleton Organization but Not for Induction of Serum Response Element Transcription

Allal¹,

Favre²,

Couderc³

et al. 2000

Journal of Biological Chemistry

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The importance of post-translational geranylgeranylation of the GTPase RhoA for its ability to induce cellular proliferation and malignant transformation is not well understood. In this manuscript we demonstrate that geranylgeranylation is required for the proper cellular localization of V14RhoA and for its ability to induce actin stress fiber and focal adhesion formation. Furthermore, V14RhoA geranylgeranylation was also required for suppressing p21 WAF transcription, promoting cell cycle progression and cellular proliferation. The ability of V14RhoA to induce focus formation and enhance plating efficiency and oncogenic Ras anchoragedependent growth was also dependent on its geranylgeranylation. The only biological activity of V14RhoA that was not dependent on its prenylation was its ability to induce serum response element transcriptional activity. Furthermore, we demonstrate that a farnesylated form of V14RhoA was also able to bind RhoGDI-1, was able to induce cytoskeleton organization, proliferation, and transformation, and was just as potent as geranylgeranylated V14RhoA at suppressing p21 WAF transcriptional activity. These results demonstrate that RhoA geranylgeranylation is required for its biological activity and that the nature of the lipid modification is not critical.Small G proteins of the Ras superfamily are regulatory proteins whose activity is controlled by a GDP/GTP cycle. Several members of the Ras superfamily are regulators of signaling pathways that control cell growth, differentiation, and oncogenic transformation as well as actin cytoskeletal organization (1). The Rho protein branch of this superfamily includes at least eight distinct Rho families (RhoA, B, C, D, and G, Rac1 and 2, TC10, Cdc42, and Rnd1, 2, and 3) (2) that are regulated by Rho-GTPase activating proteins and a large family of guanine nucleotide exchange factors of the Dbl family proteins. Moreover Rho guanine nucleotide dissociation inhibitors (RhoGDIs) 1 stabilize the inactive GDP-bound form of the Rho proteins. Rho proteins notably regulate signal transduction from cell surface receptors to intracellular molecules and are involved in a variety of cellular processes including cell morphology (3), motility (4), cytokinesis (5, 6), cell proliferation (7, 8), and tumor progression (9 -11).Ras and Rho proteins are post-translationally modified by the isoprenoid lipids, farnesyl, and geranylgeranyl (12). Two prenyltransferases, farnesyltransferase (FTase) and geranylgeranyltransferase I (GGTase I), catalyze the covalent attachment of the farnesyl and geranylgeranyl groups, respectively, to the carboxyl-terminal cysteine of proteins ending in a CAAX motif (C is a cysteine, A usually aliphatic amino acid, and X any amino acid). FTase prefers CAAX sequences where X is a serine, methionine, cysteine, alanine, or glutamine, as in Ras or in nuclear lamins (13-15). When X is a leucine or isoleucine the protein, as in the Rho/Rac family of proteins, is geranylgeranylated by GGTase I (16, 17). Protein prenylation is important in target...

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“…4B). Calculations (see "Experimental Procedures") of the dissociation constants (K d ) revealed that half-maximal binding of PDE␦ to farnesyl is at 0.70 Ϯ 0.27 M, whereas that of PDE␦ to geranylgeranyl is at 19.06 Ϯ 2.41 M. The binding constant of farnesyl to PDE␦ is higher than that of another prenyl-binding protein, RhoGDI (K d ϭ 4.8 M), whereas geranylgeranyl has approximately 10 times less affinity for PDE␦ compared with RhoGDI (K d ϭ 1.6 M) (26). DISCUSSION We show that PDE␦ can interact with both farnesyl (C15) and geranylgeranyl (C20) side chains in the absence of polypeptides.…”

Section: Figmentioning

confidence: 95%

Photoreceptor cGMP Phosphodiesterase δ Subunit (PDEδ) Functions as a Prenyl-binding Protein

Zhang

Liu

Zhang

et al. 2004

Journal of Biological Chemistry

126

122

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Bovine PDE␦ was originally copurified with rod cGMP phosphodiesterase (PDE) and shown to interact with prenylated, carboxymethylated C-terminal Cys residues. Other studies showed that PDE␦ can interact with several small GTPases including Rab13, Ras, Rap, and Rho6, all of which are prenylated, as well as the Nterminal portion of retinitis pigmentosa GTPase regulator and Arl2/Arl3, which are not prenylated. We show by immunocytochemistry with a PDE␦-specific antibody that PDE␦ is present in rods and cones. We find by yeast two-hybrid screening with a PDE␦ bait that it can interact with farnesylated rhodopsin kinase (GRK1) and that prenylation is essential for this interaction. In vitro binding assays indicate that both recombinant farnesylated GRK1 and geranylgeranylated GRK7 co-precipitate with a glutathione S-transferase-PDE␦ fusion protein. Using fluorescence resonance energy transfer techniques exploiting the intrinsic tryptophan fluorescence of PDE␦ and dansylated prenyl cysteines as fluorescent ligands, we show that PDE␦ specifically binds geranylgeranyl and farnesyl moieties with a K d of 19.06 and 0.70 M, respectively. Our experiments establish that PDE␦ functions as a prenyl-binding protein interacting with multiple prenylated proteins.

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The Specific Binding of Small Molecule Isoprenoids to rhoGDP Dissociation Inhibitor (rhoGDI)

Cited by 24 publications

References 41 publications

Structural basis of recognition of farnesylated and methylated KRAS4b by PDEδ

Structural basis of recognition of farnesylated and methylated KRAS4b by PDEδ

RhoA Prenylation Is Required for Promotion of Cell Growth and Transformation and Cytoskeleton Organization but Not for Induction of Serum Response Element Transcription

Photoreceptor cGMP Phosphodiesterase δ Subunit (PDEδ) Functions as a Prenyl-binding Protein

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