Phosphomevalonate kinase is a cytosolic protein in humans

Hogenboom, Sietske; Tuyp, John J. M.; Espeel, Marc; Koster, Janet; Wanders, Ronald J. A.; Waterham, Hans R.

doi:10.1194/jlr.m300373-jlr200

Cited by 45 publications

(28 citation statements)

References 25 publications

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“…The punctate Xuorescence did not reXect a mitochondrial localization, as determined by co-localization studies using the mitochondrial marker cytochrome c (data not shown); a lysosomal localization, as determined by co-localization studies using the lysosome markers Limp-1 and Lamp-2 (data not shown); or endosomal localization, as determined by co-localization studies using antibodies to early endosome antigen 1 (EEA1) and Rab5 (data not shown). Interestingly, these observations are similar to those published by Hogenboom et al (2004b), who observed a punctate pattern that did not co-localize with the punctate pattern of the peroxisomal catalase or the PMP ALDP (adrenoleukodystrophy protein). Thus, they reached the conclusion that this enzyme was not localized to peroxisomes.…”

Section: Phosphomevalonate Kinasesupporting

confidence: 91%

Localization of the pre-squalene segment of the isoprenoid biosynthetic pathway in mammalian peroxisomes

Kovacs

Tape

Shackelford

et al. 2006

Histochem Cell Biol

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Previous studies have indicated that the early steps in the isoprenoid/cholesterol biosynthetic pathway occur in peroxisomes. However, the role of peroxisomes in cholesterol biosynthesis has recently been questioned in several reports concluding that three of the peroxisomal cholesterol biosynthetic enzymes, namely mevalonate kinase, phosphomevalonate kinase, and mevalonate diphosphate decarboxylase, do not localize to peroxisomes in human cells even though they contain consensus peroxisomal targeting signals. We re-investigated the subcellular localization of the cholesterol biosynthetic enzymes of the pre-squalene segment in human cells by using new stable isotopic techniques and data computations with isotopomer spectral analysis, in combination with immunoXuorescence and cell permeabilization techniques. Our present Wndings clearly show and conWrm previous studies that the pre-squalene segment of the cholesterol biosynthetic pathway is localized to peroxisomes. In addition, our data are consistent with the hypothesis that acetyl-CoA derived from peroxisomal -oxidation of very long-chain fatty acids and medium-chain dicarboxylic acids is preferentially channeled to cholesterol synthesis inside the peroxisomes without mixing with the cytosolic acetyl-CoA pool.

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Section: Phosphomevalonate Kinasesupporting

confidence: 91%

Localization of the pre-squalene segment of the isoprenoid biosynthetic pathway in mammalian peroxisomes

Kovacs

Tape

Shackelford

et al. 2006

Histochem Cell Biol

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show abstract

“…These results have been challenged by data supporting the possibility that the biosynthetic reactions leading to mammalian sterols (e.g. cholesterol) occur mostly in the cytosol (Hogenboom et al, 2004;Wanders and Waterham, 2006). Localization of the rate-limiting enzyme, HMGR, and the first committed enzyme of sterol biosynthesis, squalene synthase, to the ER are in general agreement.…”

mentioning

confidence: 61%

Peroxisomal Localization of Arabidopsis Isopentenyl Diphosphate Isomerases Suggests That Part of the Plant Isoprenoid Mevalonic Acid Pathway Is Compartmentalized to Peroxisomes

et al. 2008

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“…This enzyme was first believed to be localized in the cytosol of plants (24) and animals but later was described as mitochondrial (13) and plastidic (49). Moreover, it has been described that peroxisomes are the major site of the synthesis of FPPS from mevalonate in human cells (2), while other studies maintain a cytosolic location for the conversion of mevalonic acid to isopentenyl diphosphate (21,22). Digitonin permeabilization is a classical way to assess the subcellular localization of a protein.…”

Section: Resultsmentioning

confidence: 99%

Farnesyl Diphosphate Synthase Is a Cytosolic Enzyme inLeishmania majorPromastigotes and Its Overexpression Confers Resistance to Risedronate

et al. 2006

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Farnesyl diphosphate synthase is the most likely molecular target of aminobisphosphonates (e.g., risedronate), a set of compounds that have been shown to have antiprotozoal activity both in vitro and in vivo. This protein, together with other enzymes involved in isoprenoid biosynthesis, is an attractive drug target, yet little is known about the compartmentalization of the biosynthetic pathway. Here we show the intracellular localization of the enzyme in wild-type Leishmania major promastigote cells and in transfectants overexpressing farnesyl diphosphate synthase by using purified antibodies generated towards a homogenous recombinant Leishmania major farnesyl diphosphate synthase protein. Indirect immunofluorescence, together with immunoelectron microscopy, indicated that the enzyme is mainly located in the cytoplasm of both wild-type cells and transfectants. Digitonin titration experiments also confirmed this observation. Hence, while the initial step of isoprenoid biosynthesis catalyzed by 3-hydroxy-3-methylglutaryl-coenzyme A reductase is located in the mitochondrion, synthesis of farnesyl diphosphate by farnesyl diphosphate synthase is a cytosolic process. Leishmania major promastigote transfectants overexpressing farnesyl diphosphate synthase were highly resistant to risedronate, and the degree of resistance correlated with the increase in enzyme activity. Likewise, when resistance was induced by stepwise selection with the drug, the resulting resistant promastigotes exhibited increased levels of farnesyl diphosphate synthase. The overproduction of protein under different conditions of exposure to risedronate further supports the hypothesis that this enzyme is the main target of aminobisphosphonates in Leishmania cells.

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Phosphomevalonate kinase is a cytosolic protein in humans

Cited by 45 publications

References 25 publications

Localization of the pre-squalene segment of the isoprenoid biosynthetic pathway in mammalian peroxisomes

Localization of the pre-squalene segment of the isoprenoid biosynthetic pathway in mammalian peroxisomes

Peroxisomal Localization of Arabidopsis Isopentenyl Diphosphate Isomerases Suggests That Part of the Plant Isoprenoid Mevalonic Acid Pathway Is Compartmentalized to Peroxisomes

Farnesyl Diphosphate Synthase Is a Cytosolic Enzyme inLeishmania majorPromastigotes and Its Overexpression Confers Resistance to Risedronate

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