The biological role of dolichol

Chojnacki, Tadeusz; Dallner, Gustav

doi:10.1042/bj2510001

Cited by 286 publications

(195 citation statements)

References 83 publications

Supporting

Mentioning

192

Contrasting

Unclassified

Order By: Relevance

“…To validate the ability of the methods to resolve Dol-P of varying lengths, a mixed Dol-P standard comprising C85 -C105-Dol-P, which are typical for mammalian cells [1], was labelled with 9-ADM and separated by HPLC. A dimethylation and subsequent selective demethylation was included in the Dol-P preparation to improve product stability as established by Yamada et al [15].…”

Section: Resultsmentioning

confidence: 99%

“…Dolichol-phosphate, mass spectrometry, HPLC, cholesterol pathway, statin, glycosylation 3 Introductory statement Dolichols (Dol) are essential components of eukaryotic membranes, where they contribute to the organisation and fluidity of the lipid bilayer and enhance vesicle fusion [1]. Their composition varies between the kingdoms of life in respect to the number of isoprene subunits, although all Dol are assembled in the identical trans-trans-polycis conformation.…”

Section: Introductionmentioning

confidence: 99%

“…In contrast to prokaryotic polyprenols, Dol α-isoprene residues are saturated [2]. Starting from acetate, the pathway leading to the formation of Dol leads via mevalonate to farnesyl-pyrophosphate and is up to this point identical to the cholesterol and ubiquinone biosynthesis pathway [1]. The enzyme cis-prenyltransferase catalyzes the elongation of farnesyl-pyrophosphate by sequential condensation of isopentenyl-pyrophosphate [2].…”

mentioning

confidence: 99%

See 2 more Smart Citations

HPLC and mass spectrometry analysis of dolichol-phosphates at the cell culture scale

Haeuptle¹,

Hülsmeier²,

Hennet³

2010

Analytical Biochemistry

View full text Add to dashboard Cite

Dolichols (Dol) are polyprenol lipids that are essential structural components of eukaryotic membranes. In addition, the phosphorylated derivatives of Dol function as lipid anchor of mono-and oligosaccharide precursors involved in protein glycosylation.The biological importance of Dol-phosphates (Dol-P) is illustrated by the severe outcome of human disorders linked to Dol biosynthetic defects, such as Dol-kinase deficiency. For characterization of inherited human diseases and evaluation of therapeutic trials, cultured cells often serve as a sole possible source for experimentation. Limited amounts of cell culture material render the quantitative analysis of Dol a challenging task. Here, we present HPLC and mass spectrometry based approaches to analyse and quantitate Dol-P from cultured human cells. The composition of naturally occurring Dol-P and the saturation state of the α-isoprene units was identified by negative ion electrospray ionization mass spectrometry. Furthermore, fluorescently labelled Dol-P were separated by HPLC and quantified by comparison to known amounts of the internal standard polyprenol-P. The effect of pravastatin, a 3-hydroxy-3-methyl-glutaryl coenzyme-A reductase inhibitor, on the formation of Dol-P in HeLa cells was investigated. As expected, this treatment led to a decrease of Dol-P down to 35% of normal levels. KeywordsDolichol-phosphate, mass spectrometry, HPLC, cholesterol pathway, statin, glycosylation 3 Introductory statement Dolichols (Dol) are essential components of eukaryotic membranes, where they contribute to the organisation and fluidity of the lipid bilayer and enhance vesicle fusion [1]. Their composition varies between the kingdoms of life in respect to the number of isoprene subunits, although all Dol are assembled in the identical trans-trans-polycis conformation. In contrast to prokaryotic polyprenols, Dol α-isoprene residues are saturated [2]. Starting from acetate, the pathway leading to the formation of Dol leads via mevalonate to farnesyl-pyrophosphate and is up to this point identical to the cholesterol and ubiquinone biosynthesis pathway [1]. The enzyme cis-prenyltransferase catalyzes the elongation of farnesyl-pyrophosphate by sequential condensation of isopentenyl-pyrophosphate [2]. Prior to the NADPH-dependent reduction of the α-isoprene unit, the entirely unsaturated polyprenol-pyrophosphate intermediates are dephosphorylated by mono-or pyrophosphate phosphatases. In a terminatory step, the To date, only few techniques have been described for the analysis and quantification of 12;13]. HPLC-based methods separate radio-labelled or fluorescently tagged polyprenol-P and are mainly restricted to tissue-derived samples [14; 15]. Whereas several mass spectrometry (MS) methods for the analysis of free Dol have been described [16; 17; 18; 19; 20; 21], no MS method has been established to analyse Dol-P from mammalian sources, despite the essential role of Dol-P in protein glycosylation [4].Solely, Griffiths and co-workers have configured their MS setup to s...

show abstract

Section: Resultsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

mentioning

confidence: 99%

See 1 more Smart Citation

HPLC and mass spectrometry analysis of dolichol-phosphates at the cell culture scale

Haeuptle¹,

Hülsmeier²,

Hennet³

2010

Analytical Biochemistry

View full text Add to dashboard Cite

show abstract

Section: Discussionmentioning

confidence: 99%

“…2I). This was expected because the main function of dolichols, as dolichyl phosphate, is their participation in glycoprotein synthesis in the endoplasmic reticulum, where the dolichol carrier is recycled (Chojnacki & Dallner 1988, Burda & Aebi 1999.In this framework, the metabolite levels of most of the intermediates of the MEP pathway showed an increase in trophozoite and a decrease in schizont stages, when ubiquinones were accumulated. Interestingly, metabolic profiles showed that DOXP and CDP-ME were highly accumulated when compared to the other intermediates, mainly in the trophozoite and schizont stages (Fig.…”

mentioning

confidence: 99%

Effect of fosmidomycin on metabolic and transcript profiles of the methylerythritol phosphate pathway in Plasmodium falciparum

Cassera

Merino

Peres

et al. 2007

Mem. Inst. Oswaldo Cruz

View full text Add to dashboard Cite

Key words: Plasmodium falciparum -malaria -fosmidomycin -isoprenoid biosynthesis -real time polymerase chain reaction Malaria is a leading cause of morbidity and mortality in the tropical regions, with 300 to 500 million clinical cases and 1.5 to 2.7 million deaths per year (Snow et al. 2005). With the availability of the complete genome sequence from Plasmodium falciparum, increasing attention has focused on transcript profiling and proteomic analyses of the parasite stages responsible for severe disease and pathogenesis in humans (Florens et al. 2002, Bozdech et al. 2003, Le Roch et al. 2003, Nirmalan et al. 2004, Llinas et al. 2006.Two different biosynthetic routes are used to produce isopentenyl diphosphate (IPP) and dimethylallyl diphosphate (DMAPP) for isoprenoid biosynthesis including ubiquinones, dolichols and the prenylation of proteins (Sacchettini & Poulter 1997, Sinensky 2000, Barkovich & Liao 2001. In mammals, plants (cytoplasm), fungi, some bacteria and several protozoa, the isoprenic units are derived from the classical mevalonate pathway (Goldstein & Brown 1990). In plastids of plants, several algae, eubacteria, cyanobacteria and apicomplexa (apicoplast), the 2C-methyl-D-erythritol-4-phosphate (MEP) pathway produces IPP and DMAPP (Rohmer 1999). The MEP pathway starts with the condensation of pyruvate and glyceraldehyde-3-phosphate (GAP), which yields 1-deoxy-D-xylulose-5-phosphate (DOXP) catalyzed by DOXP synthase (DXS); for reviews see references (Lichtenthaler 1999, Rohmer 1999, Eisenreich et al. 2004, Rodriguez-Concepcion 2004. DOXP reductoisomerase (DXR) then catalyzes the intramolecular rearrangement and reduction of DOXP to MEP. The activity of this enzyme is specifically inhibited by fosmidomycin (Kuzuyama et al. 1998). Several subsequent reaction steps are necessary for the conversion of MEP to IPP and DMAPP (Fig. 1).Discovery of the MEP pathway for isoprenoid biosynthesis in P. falciparum revealed several antimalarial drug targets (Jomaa et al. 1999). Jomaa and co-workers demonstrated that fosmidomycin and its derivate FR900098, are able to inhibit the growth of P. falciparum in culture and to cure mice infected with the related malaria parasite, P. vinckei (Jomaa et al. 1999). Recent field trials in humans have also demonstrated the effectiveness of fosmidomycin in the treatment of human malarial infections, but it has to be administered for more than four days when used alone (Missinou et al. 2002, Borrmann et al. 2005. Recently, biochemical and mass spectrometric analyses revealed that the MEP pathway is functionally active in all intraerythrocytic stages of P. falciparum (Cassera et al. 2004).In this study we characterized the effect of fosmidomycin on the metabolic levels of each intermediate of the MEP pathway as well as dolichol and ubiquino- nes in ring, trophozoite and schizont stages of P. falciparum parasites and correlated these to the steadystate MEP enzyme transcript levels under drug pressure. MATERIALS AND METHODSExperimental design -Three characteristic developmental sta...

show abstract

Content of dolichol and retinol in isolated rat non‐parenchymal liver cells

Nanni

Canepa

Majorani

et al. 1998

Cell Biochemistry & Function

View full text Add to dashboard Cite

The liver sinusoids, that are considered as a functional unit, harbour four types of sinusoidal cells (Ito, Kupffer, endothelial and pit cells). Dolichol content has been determined in many tissues and subcellular compartments, alteration has been reported in many types of liver injury, but until now no data are available on its content in every type of sinusoidal non-parenchymal liver cells. Dolichol and retinol metabolism might intersect in their traffic in biological membranes. Intercellular as well as intracellular exchange of retinoids is an essential element of important processes occurring in liver cells. It has been suggested that the role of dolichol, besides being a carrier of oligosaccharides in the biosynthesis of N-linked glycoproteins, may be to modify membrane fluidity and permeability, and facilitate fusion of membranes. Dolichol in the membrane is intercalated between the two-halves of the phospholipid bilayer, but its exact disposition is not known and the movement and distribution of retinoid in membranes may vary with the geometry of the membranes. Therefore the aim of this study is to obtain a global understanding of the sinusoidal system regarding dolichol and retinol content in each type of isolated rat liver sinusoidal cell, in normal conditions and after vitamin A administration. The information that can be drawn from the present results is that with normal vitamin A status of the animal, the dolichol content is almost uniform in all liver cells. After vitamin A supplementation, a great increase of dolichol, together with the known increase of retinol, can be measured only in a subpopulation of the Ito cells, the Ito-1 subfraction. Therefore in the cells that are present in the hepatic sinusoid, different pools of dolichol may have separate functions. Because retinol traffic among cells, membranes and plasma still remains to be fully understood, roles of dolichol in the exchange of vitamin A among sinusoidal liver cells are discussed.

show abstract

The biological role of dolichol

Cited by 286 publications

References 83 publications

HPLC and mass spectrometry analysis of dolichol-phosphates at the cell culture scale

HPLC and mass spectrometry analysis of dolichol-phosphates at the cell culture scale

Effect of fosmidomycin on metabolic and transcript profiles of the methylerythritol phosphate pathway in Plasmodium falciparum

Content of dolichol and retinol in isolated rat non‐parenchymal liver cells

Contact Info

Product

Resources

About