RhoB prenylation is driven by the three carboxyl-terminal amino acids of the protein: Evidenced in vivo by an anti-farnesyl cysteine antibody (original) (raw)
Related papers
Protein prenylation: a pivotal posttranslational process
Biochemical and Biophysical Research Communications, 2003
The joining of the 15-carbon farnesyl group ðC 15 H 25 Þ and the 20-carbon geranylgeranyl group ðC 20 H 33 Þ to protein-cysteines at or near their carboxy-termini is catalyzed by protein farnesyltransferase (FTase) and protein geranylgeranyltransferase-I and II (GGTase-I and GGTase-II) . The prenyltransferases are heterodimers consisting of a-and b-subunits with combined molecular masses ranging from 91 to 98 kDa. The asubunits of FTase and GGTase-I are the same, and the b-subunits differ. The b-subunits of the three enzymes are homologous to the a-subunits and to each other. The overall reactions are shown by the following chemical equations:
Breakthroughs and Views Protein prenylation: a pivotal posttranslational processq
The joining of the 15-carbon farnesyl group ðC 15 H 25 Þ and the 20-carbon geranylgeranyl group ðC 20 H 33 Þ to protein-cysteines at or near their carboxy-termini is catalyzed by protein farnesyltransferase (FTase) and protein geranylgeranyltransferase-I and II (GGTase-I and GGTase-II) . The prenyltransferases are heterodimers consisting of a-and b-subunits with combined molecular masses ranging from 91 to 98 kDa. The asubunits of FTase and GGTase-I are the same, and the b-subunits differ. The b-subunits of the three enzymes are homologous to the a-subunits and to each other. The overall reactions are shown by the following chemical equations:
Genome Biology, 2003
Three different protein prenyltransferases (farnesyltransferase and geranylgeranyltransferases I and II) catalyze the attachment of prenyl lipid anchors 15 or 20 carbons long to the carboxyl termini of a variety of eukaryotic proteins. Farnesyltransferase and geranylgeranyltransferase I both recognize a 'Ca 1 a 2 X' motif on their protein substrates; geranylgeranyltransferase II recognizes a different, non-CaaX motif. Each enzyme has two subunits. The genes encoding CaaX protein prenyltransferases are considerably longer than those encoding non-CaaX subunits, as a result of longer introns. Alternative splice forms are predicted to occur, but the extent to which each splice form is translated and the functions of the different resulting isoforms remain to be established. Farnesyltransferase-inhibitor drugs have been developed as anti-cancer agents and may also be able to treat several other diseases. The effects of these inhibitors are complicated, however, by the overlapping substrate specificities of geranylgeranyltransferase I and farnesyltransferase.
BMC biochemistry, 2006
Available in vitro and in vivo methods for verifying protein substrates for posttranslational modifications via farnesylation or geranylgeranylation (for example, autoradiography with 3H-labeled anchor precursors) are time consuming (weeks/months), laborious and suffer from low sensitivity. We describe a new technique for detecting prenyl anchors in N-terminally glutathione S-transferase (GST)-labeled constructs of target proteins expressed in vitro in rabbit reticulocyte lysate and incubated with 3H-labeled anchor precursors. Alternatively, hemagglutinin (HA)-labeled constructs expressed in vivo (in cell culture) can be used. For registration of the radioactive marker, we propose to use a thin layer chromatography (TLC) analyzer. As a control, the protein yield is tested by Western blotting with anti-GST- (or anti-HA-) antibodies on the same membrane that has been previously used for TLC-scanning. These protocols have been tested with Rap2A, v-Ki-Ras2 and RhoA (variant RhoA63L) inc...
Expression Cloning of a Novel Farnesylated Protein, RDJ2, Encoding a DnaJ Protein Homologue
Archives of Biochemistry and Biophysics, 1997
isoprenoids, either 15-carbon farnesyl or 20-carbon ger-The CAAX farnesyltransferase is a heterodimeric en-anylgeranyl, can be found attached to proteins in thiozyme that attaches a farnesyl group to a single cysteine ether linkage to conserved cysteine residues at or near in cellular proteins which terminate in the sequence their carboxyl terminus. Most prenylated proteins are CAAX, where C is cysteine, A is an aliphatic amino acid, thought to serve as regulators of signal transduction and X is most often methionine or serine. Substrates and membrane trafficking, and prenylation has been include the p21 ras proteins, nuclear lamins, and a series shown to promote both the protein-protein and proof retinal proteins. To date, a limited number of subtein-membrane interactions of these molecules (2, 3). strates for the farnesyltransferase have been identified, Prenylation provides a mechanism for the membrane predominantly by demonstration of the attachment of localization of proteins which lack a transmembrane a farnesyl group to previously identified cDNA clones domain and appears to be a prerequisite for their in which encode proteins containing an appropriate carvivo activity.
The FASEB Journal
Isoprenylation is a posttranslational modification that involves the formation of thioether bonds between cysteine and isoprenyl groups derived from pyrophosphate intermediates of the cholesterol biosynthetic pathway. Numerous isoprenylated proteins have been detected in mammalian cells. Those identified include K-, N-, and H-p21, ras-related GTP-binding proteins such as G25K (G), nuclear lamin B and prelamin A, and the 'y subunits of heterotrimeric G proteins. The modified cysteine is located in the fourth position from the carboxyl terminus in every protein where this has been studied. For p2l, the last three amino I isopentlny-Pe Kin,,. Kiness D.csrboxyl.se isom.rso. Isop.nt.nyl-tRNA 4-DIm.thylallyl-PP DIm.thylallyl-PP Olmsthyisilyl Ad.nOsln. Trsnstsrss.
Monatshefte für Chemie - Chemical Monthly, 2006
Since 1979, when prenylation has been first discovered as chemical oddity of a yeast mating factor, the two forms of this posttranslational modification of proteins (farnesylation and geranylgeranylation) have been found as wide spread among proteins from Eukarya and their viruses. This review attempts to summarize as comprehensively as possible the enzymological processes of prenylation and the various aspects of their biological significance. The substrate proteins of prenyltransferases are known to carry a sequence signal composed of a cysteine-containing 4-5 residue stretch at the utmost C-terminal end that is N-terminally preceded by a flexible and polar linker region of ca. 10 residues. Postprenylation processing of substrate proteins can involve C-terminal proteolysis, C-terminal carboxyl methylation, and other steps of maturation. The prenyl anchor functions as module for membrane attachment or for protein-protein interaction. Prenyl anchor carrying proteins fulfill a large array of functions in signaling and regulation of cellular processes. Therefore, they are involved in the pathogenesis of a variety of human diseases, the most prominent one being cancer. Farnesyltransferase inhibitors show surprisingly high efficiency in controlling tumor growth in model systems but, so far, clinical trials with human patients have remained without the desired success. Interference into prenylation pathways appears also a promising treatment principle in a variety of parasitic diseases.