Decreased expression of prenyl diphosphate synthase subunit 2 correlates with reduced survival of patients with gastric cancer

ABSTRACT Mycobacterium smegmatis has been shown to contain two forms of polyprenyl phosphate (Pol-P), while Mycobacterium tuberculosis contains only one. Utilizing subcellular fractions from M. smegmatis and M. tuberculosis, we show that Pol-P synthesis is different in these species. The specific activities of the prenyl diphosphate synthases in M. tuberculosis are 10- to 100-fold lower than those in M. smegmatis. In M. smegmatis decaprenyl diphosphate and heptaprenyl diphosphate were the main products synthesized in vitro, whereas in M. tuberculosis only decaprenyl diphosphate was synthesized. The data from both organisms suggest that geranyl diphosphate is the allylic substrate for two distinct prenyl diphosphate synthases, one located in the cell membrane that synthesizes ω,E,Z-farnesyl diphosphate and the other present in the cytosol that synthesizes ω,E,E,E-geranylgeranyl diphosphate. In M. smegmatis, the ω,E,Z-farnesyl diphosphate is utilized by a membrane-associated prenyl diphosphate synthase activity to generate decaprenyl diphosphate, and the ω,E,E,E-geranylgeranyl diphosphate is utilized by a membrane-associated activity for the synthesis of the heptaprenyl diphosphate. In M. tuberculosis, however, ω,E,E,E-geranylgeranyl diphosphate is not utilized for the synthesis of heptaprenyl diphosphate. Thus, the difference in the compositions of the Pol-P ofM. smegmatis and M. tuberculosis can be attributed to distinct enzymatic differences between these two organisms.

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Decaprenyl Diphosphate Synthesis in Mycobacterium tuberculosis

Journal of Bacteriology ◽

10.1128/jb.186.22.7564-7570.2004 ◽

2004 ◽

Vol 186 (22) ◽

pp. 7564-7570 ◽

Cited By ~ 43

Author(s):

Devinder Kaur ◽

Patrick J. Brennan ◽

Dean C. Crick

Keyword(s):

Mycobacterium Tuberculosis ◽

Divalent Cations ◽

Expression System ◽

Catalytic Efficiency ◽

Farnesyl Diphosphate ◽

Isopentenyl Diphosphate ◽

Prenyl Diphosphate ◽

Mycobacterial Cell Wall ◽

Prenyl Diphosphate Synthase

ABSTRACT Z-prenyl diphosphate synthases catalyze the sequential condensation of isopentenyl diphosphate with allylic diphosphates to synthesize polyprenyl diphosphates. In mycobacteria, these are precursors of decaprenyl phosphate, a molecule which plays a central role in the biosynthesis of essential mycobacterial cell wall components, such as the mycolyl-arabinogalactan-peptidoglycan complex and lipoarabinomannan. Recently, it was demonstrated that open reading frame Rv2361c of the Mycobacterium tuberculosis H37Rv genome encodes a unique prenyl diphosphate synthase (M. C. Schulbach, P. J. Brennan, and D. C. Crick, J. Biol. Chem. 275:22876-22881, 2000). We have now purified the enzyme to near homogeneity by using an Escherichia coli expression system and have shown that the product of this enzyme is decaprenyl diphosphate. Rv2361c has an absolute requirement for divalent cations and an optimal pH range of 7.5 to 8.5, and the activity is stimulated by both detergent and dithiothreitol. The enzyme catalyzes the addition of isopentenyl diphosphate to geranyl diphosphate, neryl diphosphate, ω,E,E-farnesyl diphosphate, ω,E,Z-farnesyl diphosphate, or ω,E,E,E-geranylgeranyl diphosphate, with Km values for the allylic substrates of 490, 29, 84, 290, and 40 μM, respectively. The Km value for isopentenyl diphosphate is 89 μM. The catalytic efficiency is greatest when ω,E,Z-farnesyl diphosphate is used as the allylic acceptor, suggesting that this is the natural substrate in vivo, a conclusion that is supported by previous structural studies of decaprenyl phosphoryl mannose isolated from M. tuberculosis. This is the first report of a bacterial Z-prenyl diphosphate synthase that preferentially utilizes an allylic diphosphate primer having the α-isoprene unit in the Z configuration, indicating that Rv1086 (ω,E,Z-farnesyl diphosphate synthase) and Rv2361c act sequentially in the biosynthetic pathway that leads to the formation of decaprenyl phosphate in M. tuberculosis.

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Faculty Opinions recommendation of The biosynthetic origin of irregular monoterpenes in Lavandula: isolation and biochemical characterization of a novel cis-prenyl diphosphate synthase gene, lavandulyl diphosphate synthase.

Faculty Opinions – Post-Publication Peer Review of the Biomedical Literature ◽

10.3410/f.718001912.793475320 ◽

2013 ◽

Author(s):

Eran Pichersky

Keyword(s):

Biochemical Characterization ◽

Prenyl Diphosphate ◽

Prenyl Diphosphate Synthase

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Gene network reconstruction identifies the authentic trans ‐prenyl diphosphate synthase that makes the solanesyl moiety of ubiquinone‐9 in Arabidopsis

The Plant Journal ◽

10.1111/j.1365-313x.2011.04796.x ◽

2011 ◽

Vol 69 (2) ◽

pp. 366-375 ◽

Cited By ~ 31

Author(s):

Anne‐Lise Ducluzeau ◽

Yashitola Wamboldt ◽

Christian G. Elowsky ◽

Sally A. Mackenzie ◽

Robert C. Schuurink ◽

...

Keyword(s):

Gene Network ◽

Network Reconstruction ◽

Gene Network Reconstruction ◽

Prenyl Diphosphate ◽

Prenyl Diphosphate Synthase

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Novel Medium-Chain Prenyl Diphosphate Synthase from the Thermoacidophilic Archaeon Sulfolobus solfataricus

Journal of Bacteriology ◽

10.1128/jb.184.3.615-620.2002 ◽

2002 ◽

Vol 184 (3) ◽

pp. 615-620 ◽

Cited By ~ 21

Author(s):

Hisashi Hemmi ◽

Satoru Ikejiri ◽

Satoshi Yamashita ◽

Tokuzo Nishino

Keyword(s):

Sulfolobus Solfataricus ◽

Biological Significance ◽

Open Reading Frames ◽

The Other ◽

Whole Genome Sequence ◽

Geranylgeranyl Diphosphate ◽

Product Specificity ◽

Medium Chain ◽

Prenyl Diphosphate ◽

Prenyl Diphosphate Synthase

ABSTRACT Two open reading frames which encode the homologues of (all-E) prenyl diphosphate synthase are found in the whole-genome sequence of Sulfolobus solfataricus, a thermoacidophilic archaeon. It has been suggested that one is a geranylgeranyl diphosphate synthase gene, but the specificity and biological significance of the enzyme encoded by the other have remained unclear. Thus, we isolated the latter by the PCR method, expressed the enzyme in Escherichia coli cells, purified it, and characterized it. The archaeal enzyme, 281 amino acids long, is highly thermostable and requires Mg2+ and Triton X-100 for full activity. It catalyzes consecutive E-type condensations of isopentenyl diphosphate with an allylic substrate such as geranylgeranyl diphosphate and yields the medium-chain product hexaprenyl diphosphate. Despite such product specificity, phylogenetic analysis revealed that the archaeal medium-chain prenyl diphosphate synthase is distantly related to the other medium- and long-chain enzymes but is closely related to eucaryal short-chain enzymes.

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