Characterization of a lysine-to-glutamic acid mutation in a conservative sequence of farnesyl diphosphate synthase from Saccharomyces cerevisiae

ABSTRACTSchistosomiasis affects over 200 million people worldwide, with over 200,000 deaths annually. Currently, praziquantel is the only drug available against schistosomiasis. We report here thatSchistosoma mansonifarnesyl diphosphate synthase (SmFPPS) and geranylgeranyl diphosphate synthase (SmGGPPS) are potential drug targets for the treatment of schistosomiasis. We expressed active, recombinantSmFPPS andSmGGPPS for subsequent kinetic characterization and testing against a variety of bisphosphonate inhibitors. RecombinantSmFPPS was found to be a soluble 44.2-kDa protein, whileSmGGPPS was a soluble 38.3-kDa protein. Characterization of the substrate utilization of the two enzymes indicates that they have overlapping substrate specificities. AgainstSmFPPS, several bisphosphonates had 50% inhibitory concentrations (IC50s) in the low micromolar to nanomolar range; these inhibitors had significantly less activity againstSmGGPPS. Several lipophilic bisphosphonates were active againstex vivoadult worms, with worm death occurring over 4 to 6 days. These results indicate that FPPS and GGPPS could be of interest in the context of the emerging resistance to praziquantel in schistosomiasis therapy.

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Enhanced (S )-linalool production by fusion expression of farnesyl diphosphate synthase and linalool synthase in Saccharomyces cerevisiae

Journal of Applied Microbiology ◽

10.1111/jam.13105 ◽

2016 ◽

Vol 121 (1) ◽

pp. 187-195 ◽

Cited By ~ 29

Author(s):

Yu Deng ◽

Mingxue Sun ◽

Sha Xu ◽

Jingwen Zhou

Keyword(s):

Saccharomyces Cerevisiae ◽

Farnesyl Diphosphate ◽

Fusion Expression ◽

Farnesyl Diphosphate Synthase ◽

Linalool Synthase

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Structural characterization of a lepidopteran type-II farnesyl diphosphate synthase from the spruce budworm, Choristoneura fumiferana : Implications for inhibitor design

Insect Biochemistry and Molecular Biology ◽

10.1016/j.ibmb.2017.11.011 ◽

2018 ◽

Vol 92 ◽

pp. 84-92 ◽

Cited By ~ 1

Author(s):

Marie-Ève Picard ◽

Audrey Nisole ◽

Catherine Béliveau ◽

Stephanie Sen ◽

Aline Barbar ◽

...

Keyword(s):

Structural Characterization ◽

Choristoneura Fumiferana ◽

Spruce Budworm ◽

Farnesyl Diphosphate ◽

Farnesyl Diphosphate Synthase ◽

Type Ii ◽

Inhibitor Design

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Functional relationships between the Saccharomyces cerevisiae cis-prenyltransferases required for dolichol biosynthesis.

Acta Biochimica Polonica ◽

10.18388/abp.2005_3511 ◽

2005 ◽

Vol 52 (1) ◽

pp. 221-232 ◽

Cited By ~ 10

Author(s):

Kariona Grabińska ◽

Grazyna Sosińska ◽

Jacek Orłowski ◽

Ewa Swiezewska ◽

Thierry Berges ◽

...

Keyword(s):

Saccharomyces Cerevisiae ◽

Enzymatic Activity ◽

Farnesyl Diphosphate ◽

Farnesyl Diphosphate Synthase ◽

Gene Encoding ◽

Yeast Saccharomyces Cerevisiae ◽

Functional Relationships ◽

Dolichyl Phosphate ◽

Preferential Synthesis

In the yeast Saccharomyces cerevisiae the RER2 and SRT1 genes encode Rer2 and Srt1 proteins with cis-prenyltransferase (cis-PT-ase) activity. Both cis-PT-ases utilize farnesyl diphosphate (FPP) as a starter for polyprenyl diphosphate (dolichol backbone) formation. The products of the Rer2 and Srt1 proteins consist of 14-17 and 18-23 isoprene units, respectively. In this work we demonstrate that deletion or overexpression of SRT1 up-regulates the activity of Rer2p and dolichol content. However, upon overexpression of SRT1, preferential synthesis of longer-chain dolichols and a decrease in the amount of the shorter species are observed. Furthermore, overexpression of the ERG20 gene (encoding farnesyl diphosphate synthase, Erg20p) induces transcription of SRT1 mRNA and increases the levels of mRNA for RER2 and DPM1 (dolichyl phosphate mannose synthase, Dpm1p). Subsequently the enzymatic activity of Rer2p and dolichol content are also increased. However, the amount of Dpm1p or its enzymatic activity remain unchanged.

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