The basis for K-Ras4B binding specificity to protein farnesyl-transferase revealed by 2 Å resolution ternary complex structures

Long, Stephen B.; Casey, Patrick J.; Beese, L.S.

doi:10.1016/s0969-2126(00)00096-4

Cited by 125 publications

(229 citation statements)

References 47 publications

Supporting

Mentioning

213

Contrasting

Order By: Relevance

“…All three enzymes in the protein prenyltransferase family require a divalent zinc ion for catalysis, and the amino acids that coordinate zinc are strictly conserved. This conservation, along with a variety of biochemical and structural data, suggests that these enzymes share a common catalytic mechanism where the zinc ion coordinates the sulfur of the peptide substrate (12,26,31,35,50,51). This interaction enhances catalysis by lowering the pK a of the peptide thiol, as well as positioning and, perhaps, activating the thiolate for nucleophilic attack on the isoprenoid diphosphate (26,31,51).…”

Section: Discussionmentioning

confidence: 95%

“…Structure of Active Ternary Complex-The crystal structures of FTase and GGTase I indicate that the substrates bound in their respective ternary and product complexes have a similar overall conformation (12,50,52). The first three isoprene units of 3-aza-GGPP are arranged along a straight line in the central cavity of the ␤-subunit of GGTase I, with the first isoprene unit shifted 1 Å relative to the crystal structure of an FPP analog bound to FTase (12).…”

Section: Discussionmentioning

confidence: 99%

See 1 more Smart Citation

Lysine β311 of Protein Geranylgeranyltransferase Type I Partially Replaces Magnesium

Hartman

Bowers

Fierke

2004

Journal of Biological Chemistry

View full text Add to dashboard Cite

Protein geranylgeranyltransferase type I (GGTase I) catalyzes the attachment of a geranylgeranyl lipid group near the carboxyl terminus of protein substrates. Unlike protein farnesyltransferase (FTase) and protein geranylgeranyltransferase type II, which require both Zn(II) and Mg(II) for maximal turnover, GGTase I turnover is dependent only on Zn(II). In FTase, the magnesium ion is coordinated by aspartate ␤352 and the diphosphate of farnesyl diphosphate to stabilize the developing charge in the transition state (Pickett, J. S., Bowers, K. E., and Fierke, C. A. Prenylation is a type of post-translational modification where a lipid group from either farnesyl diphosphate (FPP) 1 or geranylgeranyl diphosphate (GGPP) is covalently attached via a thioether linkage to a conserved cysteine residue near the carboxyl terminus of a protein (1-6; for review, see Refs. 7 and 8). The attached lipid mediates membrane association and specific protein-protein interactions, which are obligatory for proper functioning of the modified protein (9, 10). Protein farnesyltransferase (FTase), protein geranylgeranyltransferase type I (GGTase I), and protein geranylgeranyltransferase type II (GGTase II) comprise the class of zinc-catalyzed sulfur alkylation enzymes known as the protein prenyltransferases.All three protein prenyltransferases are ␣/␤ heterodimers; FTase and GGTase I share common ␣-subunits and possess ␤-subunits with 25% sequence identity (8,11,12). FTase and GGTase I prenylate proteins or peptides containing carboxylterminal CaaX (C refers to a conserved cysteine residue, a refers to an aliphatic amino acid, and X generally refers to leucine or phenylalanine for GGTase I, and methionine, serine, alanine, or glutamine for FTase) sequences (13-18). The proteins Ras, RhoB, CENP-E, and CENP-F are known substrates of FTase; GGTase I modifies the ␥-subunits of most heterotrimeric G proteins as well as many Ras-related G proteins, including members of the Rac, Rap, and Rho families (9, 19 -23). In a fashion distinct from FTase and GGTase I, GGTase II catalyzes the attachment of two geranylgeranyl groups to members of the Rab family of GTPases that contain carboxylterminal sequences of XXCC, XCXC, XCCX, and CCXX (C refers to a conserved cysteine residue, and X refers to any amino acid) (24).Recently, much of the work on prenyltransferases has focused on FTase and the protein substrate, Ras, a small GTPase in the receptor tyrosine kinase signaling pathway that is a key regulator of cell division (25). 30% of all human cancers can be linked to mutations in the ras gene, which lead to constitutively activated Ras signaling (25). The post-translational attachment of a farnesyl group by FTase is required for the transforming activity of Ras oncoproteins (25). These observations prompted a search for FTase inhibitors as novel anticancer agents (25). Subsequent research demonstrated that the form of Ras most often mutated in human cancers is K-Ras4B, a protein substrate that can be farnesylated by FTase as well as geranylgeranylated by...

show abstract

Section: Discussionmentioning

confidence: 95%

Section: Discussionmentioning

confidence: 99%

Lysine β311 of Protein Geranylgeranyltransferase Type I Partially Replaces Magnesium

Hartman

Bowers

Fierke

2004

Journal of Biological Chemistry

View full text Add to dashboard Cite

show abstract

“…A poly-basic stretch of amino acids upstream of the CaaX motif can facilitate binding of proteins such as K-Ras to PFT (ref. 17 ). Several elements of Rab proteins are recognized by Rep, and only the Rab-Rep complex is acted on by PGGT-II (ref.…”

Section: Enzymatic Processing Of Prenylated Proteinsmentioning

confidence: 99%

Therapeutic intervention based on protein prenylation and associated modifications

et al. 2006

View full text Add to dashboard Cite

In eukaryotic cells, a specific set of proteins are modified by C-terminal attachment of 15-carbon farnesyl groups or 20-carbon geranylgeranyl groups that function both as anchors for fixing proteins to membranes and as molecular handles for facilitating binding of these lipidated proteins to other proteins. Additional modification of these prenylated proteins includes C-terminal proteolysis and methylation, and attachment of a 16-carbon palmitoyl group; these modifications augment membrane anchoring and alter the dynamics of movement of proteins between different cellular membrane compartments. The enzymes in the protein prenylation pathway have been isolated and characterized. Blocking protein prenylation is proving to be therapeutically useful for the treatment of certain cancers, infection by protozoan parasites and the rare genetic disease Hutchinson-Gilford progeria syndrome.Isoprenoids are lipids made of five-carbon blocks, and they constitute one of the main classes of natural products. A subset of isoprenoids called prenyl groups are found on a variety of biological substances, including proteins. Prenylated proteins and peptides are found in most (if not all) eukaryotes. They arise from the post-translational attachment of 15-carbon farnesyl or 20-carbon geranylgeranyl groups to the C-terminal segment of proteins. Protein prenyl groups not only are hydrophobic elements that bind proteins to membranes, but, at least in some cases, they also function as molecular handles that bind to hydrophobic grooves on the surface of soluble protein factors; these factors then remove the prenylated protein from the membrane in a regulated manner. NIH Public Access Author ManuscriptNat Chem Biol. Author manuscript; available in PMC 2010 June 28. Published in final edited form as:Nat Chem Biol. 2006 October ; 2(10): 518-528. doi:10.1038/nchembio818. NIH-PA Author ManuscriptNIH-PA Author Manuscript NIH-PA Author ManuscriptProtein prenylation has been vigorously studied over the past ~15 years because it is found on several signaling proteins (including heterotrimeric G proteins) that connect cell surface receptors to intracellular effectors, and also on Ras proteins, one of the most common oncoproteins found in human tumors. Ras proteins serve as molecular switches at cellular membranes to control propagation of growth signals from cell surface receptors to nuclear transcription factors. Because Ras mutations are important in many human cancers, there has been extensive focus on Ras prenylation. Discovery of protein prenyl groups and structural varietiesThe first reports of prenylated proteins and peptides described the secreted pheromone peptides from jelly fungi 1,2 , whose structure resembles that of the well-known a-factor mating pheromone from baker's yeast (Saccharomyces cerevisiae), which contains a cysteine methylester farnesylated at the C terminus 3 . In the 1980s, studies on the timing of cholesterol biosynthesis with respect to the cell cycle in human cells led to the discovery that a compound deriv...

show abstract

“…X-ray crystallography of the FTase ternary complex in the presence of manganese showed a manganese ion bound near the diphosphate-like moiety of a FPP analog (26). Also, the FTase-catalyzed farnesylation using farnesylmonophosphate (FMP) as a substrate is not activated by magnesium ions, indicating that the diphosphate moiety is required for magnesium-induced rate acceleration (21).…”

mentioning

confidence: 99%

“…In the crystal structure of an inactive FTase ternary complex with bound manganese, the only manganese ligands observed were the two oxygens of the diphosphate-like moiety of the FPP analog (26). In most enzymes that use a catalytic magnesium ion, there is at least one carboxylate ligand from the enzyme that coordinates the magnesium ion in the active site (29,30).…”

mentioning

confidence: 99%

Mutagenesis Studies of Protein Farnesyltransferase Implicate Aspartate β352 as a Magnesium Ligand

Pickett

Bowers

Fierke

2003

Journal of Biological Chemistry

View full text Add to dashboard Cite

Protein farnesyltransferase (FTase) 1 catalyzes the transfer of a 15-carbon farnesyl chain from farnesyl diphosphate (FPP) to a C-terminal cysteine sulfur of protein substrates in the cell (1). Protein or peptide substrates of FTase contain a C-terminal CaaX sequence, whereas C represents the cysteine that becomes farnesylated, a 1 and a 2 are typically small aliphatic amino acids, and X is commonly methionine, serine, glutamine, or alanine (2). FTase is known to modify several important proteins in the cell cycle machinery, including Ras, Rho B, CENP-E, and CENP-F (3). Farnesylation of proteins enhances membrane association and specific protein-protein interactions and therefore is thought to be important for localization of proteins in the cell (4 -6). After farnesylation by FTase, proteins are further processed by the CaaX protease Rce1 and the methyltransferase Icmt, producing a Cterminal farnesylated cysteine methyl ester as the end product of this pathway (7,8). Protein farnesyltransferase inhibitors are currently being evaluated in clinical trials for the treatment of several types of cancer, including acute myeloid leukemia, metastatic breast cancer, and non-small cell lung cancer (9, 10).Farnesylation is just one type of protein prenylation; proteins can also be modified with geranylgeranyl groups in a reaction catalyzed by geranylgeranyl transferase type I or type II (GGTase I and GGTase II) (11, 12). GGTase I also recognizes protein substrates with a C-terminal CaaX motif, but GGTase I substrates most frequently contain a leucine or phenylalanine at the X position (13, 14). FTase and GGTase I are structurally homologous heterodimers with identical ␣ subunits and differing ␤ subunits (15). Prenyltransferases contain an active site zinc ion that is required for activity; FTase and GGTase II also use a magnesium ion to facilitate catalysis (16 -19). Surprisingly, GGTase I apparently catalyzes the same prenyl transfer reaction with no dependence on magnesium ions (20).In FTase, the magnesium ion has been proposed to coordinate to the diphosphate of FPP, but the exact location of the magnesium binding site has not yet been identified (21). The bound zinc ion of FTase coordinates to the cysteine sulfur of the protein substrate, lowering the pK a to form a zincthiolate at neutral pH (22, 23). In the proposed farnesylation transition state of FTase (Scheme 1), positive charge accumulates on the C-1 of FPP, and the diphosphate leaving group develops additional negative charge (24, 25). Bound magnesium is proposed to stabilize the negative charge buildup on the diphosphate moiety and activate the leaving group (21). Although magnesium is not absolutely required for FTase catalysis, millimolar concentrations of magnesium ions enhance the rate constant for product formation by 700-fold (24).Several pieces of experimental evidence indicate that the diphosphate of FPP forms part of the magnesium binding site. X-ray crystallography of the FTase ternary complex in the presence of manganese showed a manganese ion bo...

show abstract

The basis for K-Ras4B binding specificity to protein farnesyl-transferase revealed by 2 Å resolution ternary complex structures

Cited by 125 publications

References 47 publications

Lysine β311 of Protein Geranylgeranyltransferase Type I Partially Replaces Magnesium

Lysine β311 of Protein Geranylgeranyltransferase Type I Partially Replaces Magnesium

Therapeutic intervention based on protein prenylation and associated modifications

Mutagenesis Studies of Protein Farnesyltransferase Implicate Aspartate β352 as a Magnesium Ligand

Contact Info

Product

Resources

About