farnesyl-monophosphate and farnesyl-pyrophosphate

Oligoprenyl phosphates are key metabolic intermediates for the biosynthesis of steroids, the side chain of ubiquinones, and dolichols and the posttranslational isoprenylation of proteins. Farnesyl phosphates are isoprenoid phosphates that resemble polyunsaturated fatty alcohol phosphates, which we have recently shown to be the minimal pharmacophores of lysophosphatidic acid (LPA) receptors. Here we examine whether farnesyl phosphates can interact with the cell surface and nuclear receptors for LPA. Both farnesyl phosphate and farnesyl diphosphate potently and specifically antagonized LPA-elicited intracellular Ca(2+)-mobilization mediated through the LPA(3) receptor, while causing only modest inhibition at the LPA(2) receptor and no measurable effect at the LPA(1) receptor. Farnesol also inhibited LPA(3) but was much less effective. The estimated dissociation constant of LPA(3) for farnesyl phosphate is 48+/-12 nM and 155+/-30 nM for farnesyl diphosphate. The transcription factor peroxisome proliferator-activated receptor gamma (PPARgamma) binds to and is activated by LPA and its analogs including fatty alcohol phosphates. We found that both farnesyl phosphate and diphosphate, but not farnesol, compete with the binding of the synthetic PPARgamma agonist [(3)H]rosiglitazone and activate the PPARgamma-mediated gene transcription. Farnesyl monophosphate at 1 microM, but not diphosphate, activated PPARalpha and PPARbeta/delta reporter gene expression. These results indicate new potential roles for the oligoprenyl phosphates as potential endogenous modulators of LPA targets and show that the polyisoprenoid chain is recognized by some LPA receptors.

Topics: Calcium Signaling; Cell Line; Humans; In Vitro Techniques; Kinetics; Ligands; Lysophospholipids; Peroxisome Proliferator-Activated Receptors; Polyisoprenyl Phosphates; Receptors, Lysophosphatidic Acid; Sesquiterpenes; Transfection

As farnesol may serve as a nonsterol endogenous regulator of the mevalonate pathway, the possibility that a kinase specific for its phosphorylation is present in the rat liver was investigated. In the 10,000 g supernatant of rat liver, farnesyl monophosphate was synthesized in the presence of ATP. The Km value for farnesol was 2.3 microM. Various detergents inhibited the activity of the enzyme. The farnesol kinase was present in rough and in smooth I microsomes, but not in smooth II microsomes, mitochondria, peroxisomes, Golgi, or plasma membranes. The enzyme was associated with the inner, luminal surface of the vesicles. Further analyses have demonstrated that an enzymatic mechanism exists which catalyzes the phosphorylation of farnesyl-P to farnesyl-PP. Activity of the farnesyl phosphate kinase resulted in the phosphorylation of the monophosphate by CTP but not by ATP, GTP, or UTP. This enzyme is activated by low concentrations of detergents. Treatment with proteases and chemical probes indicate that this second phosphorylation reaction probably takes place on the outer, cytoplasmic surface of microsomal vesicles. These results demonstrate that rat liver microsomes contain two enzymes for the consecutive phosphorylation of farnesol to farnesyl-PP.

Topics: Adenosine Triphosphate; Animals; Cations, Divalent; Chromatography, High Pressure Liquid; Farnesol; Kinetics; Male; Microsomes, Liver; Phosphorylation; Phosphotransferases (Alcohol Group Acceptor); Phosphotransferases (Phosphate Group Acceptor); Polyisoprenyl Phosphates; Rats; Rats, Sprague-Dawley; Sesquiterpenes

Hexaprenyl-diphosphate synthase from Micrococcus luteus B-P 26 has been shown to comprise two essential components, designated as components A and B. Treatment of the synthase with sulphydryl reagents (N-ethylmaleimide, iodoacetamide or p-chloromercuribenzoate) or arginine-specific reagents (2,3-butanedione, 1,2-cyclohexanedione or phenylglyoxal) resulted in a rapid loss of the component B activity. In contrast, component A was resistant to treatment with such reagents, retaining the initial activity almost completely. Farnesyl diphosphate, isopentenyl diphosphate, farnesyl monophosphate and inorganic pyrophosphate protected the synthase against the inactivation by N-ethylmaleimide, farnesyl diphosphate being the most effective. The presence of Mg2+ was essential for the protection by isopentenyl diphosphate and inorganic pyrophosphate. For protection of the synthase activity against the inactivation by 2,3-butanedione, the presence of farnesyl diphosphate, isopentenyl diphosphate and Mg2+ was more effective than that of the individual substrates and Mg2+. Inorganic pyrophosphate provided substantial protection. In the absence of component A, the component B activity was not protected by any substrates or its analogue. These results suggest that the catalytic site of the synthase is formed by cooperative interaction between components A and B, and that cysteine and arginine residues on component B play important roles in the synthase activity.

Topics: Aldehydes; Alkyl and Aryl Transferases; Arginine; Butanones; Chloromercuribenzoates; Cyclohexanes; Cyclohexanones; Diacetyl; Dimethylallyltranstransferase; Diphosphates; Enzyme Activation; Ethylmaleimide; Hemiterpenes; Iodoacetamide; Magnesium; Micrococcus; Organophosphorus Compounds; p-Chloromercuribenzoic Acid; Phenylglyoxal; Polyisoprenyl Phosphates; Sesquiterpenes; Sulfhydryl Reagents; Transferases