Cloning, analysis, and bacterial expression of human farnesyl pyrophosphate synthetase and its regulation in Hep G2 cells

Sheares, B T; White, Sylvia S.; Molowa, David T.; Chan, Karen; Ding, Victor D.-H.; Kroon, Paulus A.; Bostedor, Richard G.; Karkas, John D.

doi:10.1021/bi00446a025

Cited by 58 publications

(24 citation statements)

References 34 publications

(36 reference statements)

Supporting

Mentioning

Contrasting

Unclassified

Order By: Relevance

“…which is similar to the molecular masses of other FPP synthases (40, 39, and 43 kD for the yeast, rat, and plant enzymes, respectively) (Clarke et al, 1987;Anderson et al, 1989;Sheares et al, 1989;Hugueney and Camara, 1990;Teruya et al, 1990). The deduced amino acid sequence of the encoded protein has 80% identity with and 90% similarity to that of FPP synthase identified from A. thaliana (GenBank accession No.…”

supporting

confidence: 50%

A cDNA Encoding Farnesyl Pyrophosphate Synthase in White Lupin

Attucci

Aitken²,

Ibrahim³

et al. 1995

Plant Physiol.

View full text Add to dashboard Cite

supporting

confidence: 50%

A cDNA Encoding Farnesyl Pyrophosphate Synthase in White Lupin

Attucci

Aitken²,

Ibrahim³

et al. 1995

Plant Physiol.

View full text Add to dashboard Cite

“…0.3% Triton X-100 was used to permeabilize the parasites for 30 min. 3% bovine serum albumin, 1% fish gelatin, 50 mM NH 4 Cl, and 5% goat serum in PBS were used to block at room temperature for 1 h. A 1:100 dilution of the purified anti-TgFPPS antibody and a 1:200 dilution of fluorescein isothiocyanate-coupled goat anti-guinea pig secondary antibody were incubated with the parasites for 1 h each. Alternatively, mouse monoclonal antibodies raised against T. gondii rhoptry protein 1 (ROP1) a gift from Dr. J. Dubremetz (25) and rabbit polyclonal antibodies raised against the apicoplast protein acyl carrier protein (ACP) a gift from Dr. B. Striepen (26) were used in combination with anti-TgFPPS antibody in co-localization studies.…”

Section: Sds-gel Electrophoresis and Westernmentioning

confidence: 99%

“…FPP can also be condensed with an additional molecule of IPP by geranylgeranyl-diphosphate synthase (GGPPS) to form the 20-carbon isoprenoid geranylgeranyl diphosphate (GGPP), also used in protein prenylation. Genes encoding FPPS and GGPPS, also referred as short chain prenyltransferases, have been cloned from various species, including rats (2), humans (3,4), Saccharomyces cerevisiae (5), and plants (6,7). In protist parasites, the FPPS gene has been cloned from Trypanosoma cruzi (8) and Trypanosoma brucei (9).…”

mentioning

confidence: 99%

The Farnesyl-diphosphate/Geranylgeranyl-diphosphate Synthase of Toxoplasma gondii Is a Bifunctional Enzyme and a Molecular Target of Bisphosphonates

Ling

Miranda

et al. 2007

Journal of Biological Chemistry

115

View full text Add to dashboard Cite

Farnesyl-diphosphate synthase (FPPS) catalyzes the synthesis of farnesyl diphosphate, an important precursor of sterols, dolichols, ubiquinones, and prenylated proteins. We report the cloning and characterization of two Toxoplasma gondii farnesyl-diphosphate synthase (TgFPPS) homologs. A single genetic locus produces two transcripts, TgFPPS and TgFPPSi, by alternative splicing. Both isoforms were heterologously expressed in Escherichia coli, but only TgFPPS was active. The protein products predicted from the nucleotide sequences have 646 and 605 amino acids and apparent molecular masses of 69.5 and 64.5 kDa, respectively. Several conserved sequence motifs found in other prenyl-diphosphate synthases are present in both TgFPPSs. TgFPPS was also expressed in the baculovirus system and was biochemically characterized. In contrast to the FPPS of other eukaryotic organisms, TgFPPS is bifunctional, catalyzing the formation of both farnesyl diphosphate and geranylgeranyl diphosphate. TgFPPS localizes to the mitochondria, as determined by the co-localisation of the affinity-purified antibodies against the protein with MitoTracker, and in accord with the presence of an N-terminal mitochondria-targeting signal in the protein. This enzyme is an attractive target for drug development, because the order of inhibition of the enzyme by a number of bisphosphonates is the same as that for inhibition of parasite growth. In summary, we report the first bifunctional farnesyl-diphosphate/geranylgeranyldiphosphate synthase identified in eukaryotes, which, together with previous results, establishes this enzyme as a valid target for the chemotherapy of toxoplasmosis.Toxoplasma gondii is a pathogenic protozoan parasite that infects a wide range of vertebrate hosts, including humans. T. gondii has been recognized as a major opportunistic pathogen of fetuses from recently infected mothers and of immunocompromised patients, i.e. those with AIDS. Toxoplasmic encephalitis is associated with high mortality and morbidity and is one of the most common opportunistic infections of the central nervous system in human immunodeficiency virus-infected patients (1). The chemotherapy for toxoplasmosis is not ideal especially for the AIDS patient because there is relapse of the infection if the treatment is halted because of intolerance of the patient to the side effects. Almost all the drugs in use have toxicity associated with their continued use. In addition, most of the recommended treatments do not have an effect on the slow growing bradyzoite forms.Isoprenoids are the most diverse and abundant compounds occurring in nature. Many types of isoprenoids (e.g. steroids, cholesterol, retinoids, carotenoids, ubiquinones, and prenyl groups bound to proteins) are essential components of the cellular machinery of all organisms because of their roles in a variety of biological processes. Despite their structural and functional variety, all isoprenoids derive from a common precursor, isopentenyl diphosphate (IPP), 2 and its isomer, dimethylallyl diphosphate (...

show abstract

“…It catalyzes 2 consecutive condensations of isopentenyl diphosphate with allylic diphosphates, dimethylallyl diphosphate, and the resulting geranyl diphosphate into FPP, which is used in the biosynthesis of sterols, dolichols, mitochondrial electron transfer chain components, prenylated proteins, and a wide range of sesquiterpenoids including phytoalexins. Genes encoding FPPS have been cloned from many species, including rat (Clarke et al, 1987), human (Sheares et al, 1989), Saccharomyces cerevisiae (Anderson et al, 1989), Arabidopsis thaliana (Delourme et al, 1994), and Lupinus albus (Attucci et al, 1995).…”

Section: Introductionmentioning

confidence: 99%

Cloning and sequence analysis of the Blumea balsamifera DC farnesyl diphosphate synthase gene

Pang¹,

Guan²,

Wu³

et al. 2014

Genet. Mol. Res.

View full text Add to dashboard Cite

ABSTRACT. Blumea balsamifera DC is a member of the Compositae family and is frequently used as traditional Chinese medicine. Blumea balsamifera is rich in monoterpenes, which possess a variety of pharmacological activities, such as antioxidant, anti-bacteria, and antiviral activities. Farnesyl diphosphate synthase (FPS) is a key enzyme in the biosynthetic pathway of terpenes, playing an important regulatory role in plant growth, such as resistance and secondary metabolism. Based on the conserved oligo amino acid residues of published FPS genes from other higher plant species, a cDNA sequence, designated BbFPS, was isolated from B. balsamifera DC using polymerase chain reaction. The clones were an average of 1.6 kb and contained an open reading frame that predicted a polypeptide of 342 amino acids with 89.07% identity to FPS from other plants. The deduced amino acid sequence was dominated by hydrophobic regions and contained 2 highly conserved DDxxD motifs that are essential for proper functioning of FPS. Phylogenetic analysis indicated that FPS grouped with other Blumea balsamifera DC farnesyl diphosphate synthase composite families. Prediction of secondary structure and subcellular localization suggested that alpha helices made up 70% of the amino acids of the sequence.

show abstract

Cloning, analysis, and bacterial expression of human farnesyl pyrophosphate synthetase and its regulation in Hep G2 cells

Cited by 58 publications

References 34 publications

A cDNA Encoding Farnesyl Pyrophosphate Synthase in White Lupin

A cDNA Encoding Farnesyl Pyrophosphate Synthase in White Lupin

The Farnesyl-diphosphate/Geranylgeranyl-diphosphate Synthase of Toxoplasma gondii Is a Bifunctional Enzyme and a Molecular Target of Bisphosphonates

Cloning and sequence analysis of the Blumea balsamifera DC farnesyl diphosphate synthase gene

Contact Info

Product

Resources

About