Abstract
Based on topological studies mannosylphosphoryldolichol (Man-P-Dol) is synthesized on the cytoplasmic face of the RER, but functions as a mannosyl donor in Glc3Man9GlcNAc2-P-P-dolichol biosynthesis after the mannosyl-phosphoryl headgroup diffuses transversely to the luminal compartment. The transport of mannosylphosphorylcitronellol (Man-P- Cit), a water-soluble analogue of Man-P-Dol, by microsomal vesicles from mouse liver, has been investigated as a potential experimental approach to determine if a membrane protein(s) mediates the transbilayer movement of Man-P-Dol. For these studies beta-[3H]Man-P- Cit was synthesized enzymatically with a partially purified preparation of Man-P-undecaprenol synthase from Micrococcus luteus. The uptake of the radiolabeled water-soluble analogue was found to be (a) time dependent; (b) stereoselective; (c) dependent on an intact permeability barrier; (d) saturable; (e) protease-sensitive; and (f) highest in ER- enriched vesicles relative to Golgi complex-enriched vesicles and intact mitochondria. Consistent with the involvement of a membrane protein, the analogue did not enter synthetic phosphatidylcholine- liposomes. [3H]Man-P-Cit also was not transported by human erythrocytes. These results indicate that the transport of Man-P-Cit by sealed microsomal vesicles from mouse liver is mediated by a membrane protein transport system. It is possible that the same membrane protein(s) participates in the transbilayer movement of Man-P-Dol in the ER.
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- Abeijon C., Hirschberg C. B. Topography of glycosylation reactions in the endoplasmic reticulum. Trends Biochem Sci. 1992 Jan;17(1):32–36. doi: 10.1016/0968-0004(92)90424-8. [DOI] [PubMed] [Google Scholar]
- BARTLETT G. R. Phosphorus assay in column chromatography. J Biol Chem. 1959 Mar;234(3):466–468. [PubMed] [Google Scholar]
- Bell G. I., Burant C. F., Takeda J., Gould G. W. Structure and function of mammalian facilitative sugar transporters. J Biol Chem. 1993 Sep 15;268(26):19161–19164. [PubMed] [Google Scholar]
- Bishop W. R., Bell R. M. Assembly of the endoplasmic reticulum phospholipid bilayer: the phosphatidylcholine transporter. Cell. 1985 Aug;42(1):51–60. doi: 10.1016/s0092-8674(85)80100-8. [DOI] [PubMed] [Google Scholar]
- Camp L. A., Chauhan P., Farrar J. D., Lehrman M. A. Defective mannosylation of glycosylphosphatidylinositol in Lec35 Chinese hamster ovary cells. J Biol Chem. 1993 Mar 25;268(9):6721–6728. [PubMed] [Google Scholar]
- Carey D. J., Sommers L. W., Hirschberg C. B. CMP-N-acetylneuraminic acid: isolation from and penetration into mouse liver microsomes. Cell. 1980 Mar;19(3):597–605. doi: 10.1016/s0092-8674(80)80036-5. [DOI] [PubMed] [Google Scholar]
- Coleman R., Bell R. M. Evidence that biosynthesis of phosphatidylethanolamine, phosphatidylcholine, and triacylglycerol occurs on the cytoplasmic side of microsomal vesicles. J Cell Biol. 1978 Jan;76(1):245–253. doi: 10.1083/jcb.76.1.245. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Coste H., Martel M. B., Got R. Topology of glucosylceramide synthesis in Golgi membranes from porcine submaxillary glands. Biochim Biophys Acta. 1986 Jun 13;858(1):6–12. doi: 10.1016/0005-2736(86)90285-3. [DOI] [PubMed] [Google Scholar]
- DeLuca A. W., Rush J. S., Lehrman M. A., Waechter C. J. Mannolipid donor specificity of glycosylphosphatidylinositol mannosyltransferase-I (GPIMT-I) determined with an assay system utilizing mutant CHO-K1 cells. Glycobiology. 1994 Dec;4(6):909–915. doi: 10.1093/glycob/4.6.909. [DOI] [PubMed] [Google Scholar]
- Dotson S. B., Rush J. S., Ricketts A. D., Waechter C. J. Mannosylphosphoryldolichol-mediated O-mannosylation of yeast glycoproteins: stereospecificity and recognition of the alpha-isoprene unit by a purified mannosyltransferase. Arch Biochem Biophys. 1995 Feb 1;316(2):773–779. doi: 10.1006/abbi.1995.1103. [DOI] [PubMed] [Google Scholar]
- Englund P. T. The structure and biosynthesis of glycosyl phosphatidylinositol protein anchors. Annu Rev Biochem. 1993;62:121–138. doi: 10.1146/annurev.bi.62.070193.001005. [DOI] [PubMed] [Google Scholar]
- Fleischer B., Smigel M. Solubilization and properties of galactosyltransferase and sulfotransferase activities of Golgi membranes in Triton X-100. J Biol Chem. 1978 Mar 10;253(5):1632–1638. [PubMed] [Google Scholar]
- Fleischer S., Kervina M. Subcellular fractionation of rat liver. Methods Enzymol. 1974;31:6–41. doi: 10.1016/0076-6879(74)31005-1. [DOI] [PubMed] [Google Scholar]
- Futerman A. H., Pagano R. E. Determination of the intracellular sites and topology of glucosylceramide synthesis in rat liver. Biochem J. 1991 Dec 1;280(Pt 2):295–302. doi: 10.1042/bj2800295. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Hanover J. A., Lennarz W. J. The topological orientation of N,N'-diacetylchitobiosylpyrophosphoryldolichol in artificial and natural membranes. J Biol Chem. 1979 Sep 25;254(18):9237–9246. [PubMed] [Google Scholar]
- Herscovics A., Orlean P. Glycoprotein biosynthesis in yeast. FASEB J. 1993 Apr 1;7(6):540–550. doi: 10.1096/fasebj.7.6.8472892. [DOI] [PubMed] [Google Scholar]
- Hirschberg C. B., Snider M. D. Topography of glycosylation in the rough endoplasmic reticulum and Golgi apparatus. Annu Rev Biochem. 1987;56:63–87. doi: 10.1146/annurev.bi.56.070187.000431. [DOI] [PubMed] [Google Scholar]
- Jeckel D., Karrenbauer A., Burger K. N., van Meer G., Wieland F. Glucosylceramide is synthesized at the cytosolic surface of various Golgi subfractions. J Cell Biol. 1992 Apr;117(2):259–267. doi: 10.1083/jcb.117.2.259. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Kean E. L. Activation by dolichol phosphate-mannose of the biosynthesis of N-acetylglucosaminylpyrophosphoryl polyprenols by the retina. J Biol Chem. 1982 Jul 25;257(14):7952–7954. [PubMed] [Google Scholar]
- Kean E. L., Rush J. S., Waechter C. J. Activation of GlcNAc-P-P-dolichol synthesis by mannosylphosphoryldolichol is stereospecific and requires a saturated alpha-isoprene unit. Biochemistry. 1994 Aug 30;33(34):10508–10512. doi: 10.1021/bi00200a036. [DOI] [PubMed] [Google Scholar]
- Kean E. L. Stimulation by dolichol phosphate-mannose and phospholipids of the biosynthesis of N-acetylglucosaminylpyrophosphoryl dolichol. J Biol Chem. 1985 Oct 15;260(23):12561–12571. [PubMed] [Google Scholar]
- Klingenberg M. The ferricyanide method for elucidating the sidedness of membrane-bound dehydrogenases. Methods Enzymol. 1979;56:229–233. doi: 10.1016/0076-6879(79)56025-x. [DOI] [PubMed] [Google Scholar]
- Kornberg R. D., McConnell H. M. Inside-outside transitions of phospholipids in vesicle membranes. Biochemistry. 1971 Mar 30;10(7):1111–1120. doi: 10.1021/bi00783a003. [DOI] [PubMed] [Google Scholar]
- McCloskey M. A., Troy F. A. Paramagnetic isoprenoid carrier lipids. 2. Dispersion and dynamics in lipid membranes. Biochemistry. 1980 May 13;19(10):2061–2066. doi: 10.1021/bi00551a009. [DOI] [PubMed] [Google Scholar]
- Menon A. K., Mayor S., Schwarz R. T. Biosynthesis of glycosyl-phosphatidylinositol lipids in Trypanosoma brucei: involvement of mannosyl-phosphoryldolichol as the mannose donor. EMBO J. 1990 Dec;9(13):4249–4258. doi: 10.1002/j.1460-2075.1990.tb07873.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Mulford C. A., Osborn M. J. An intermediate step in translocation of lipopolysaccharide to the outer membrane of Salmonella typhimurium. Proc Natl Acad Sci U S A. 1983 Mar;80(5):1159–1163. doi: 10.1073/pnas.80.5.1159. [DOI] [PMC free article] [PubMed] [Google Scholar]
- PENNINGTON R. J. Biochemistry of dystrophic muscle. Mitochondrial succinate-tetrazolium reductase and adenosine triphosphatase. Biochem J. 1961 Sep;80:649–654. doi: 10.1042/bj0800649. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Raetz C. R. Biochemistry of endotoxins. Annu Rev Biochem. 1990;59:129–170. doi: 10.1146/annurev.bi.59.070190.001021. [DOI] [PubMed] [Google Scholar]
- Rodríguez-Vico F., Martínez-Cayuela M., García-Peregrín E., Ramírez H. A procedure for eliminating interferences in the lowry method of protein determination. Anal Biochem. 1989 Dec;183(2):275–278. doi: 10.1016/0003-2697(89)90479-x. [DOI] [PubMed] [Google Scholar]
- Rothman J. E., Dawidowicz E. A. Asymmetric exchange of vesicle phospholipids catalyzed by the phosphatidylcholine exhange protein. Measurement of inside--outside transitions. Biochemistry. 1975 Jul;14(13):2809–2816. doi: 10.1021/bi00684a004. [DOI] [PubMed] [Google Scholar]
- Rush J. S., Shelling J. G., Zingg N. S., Ray P. H., Waechter C. J. Mannosylphosphoryldolichol-mediated reactions in oligosaccharide-P-P-dolichol biosynthesis. Recognition of the saturated alpha-isoprene unit of the mannosyl donor by pig brain mannosyltransferases. J Biol Chem. 1993 Jun 25;268(18):13110–13117. [PubMed] [Google Scholar]
- Rush J. S., Waechter C. J. An anion-exchange radioassay for glucose 6-phosphate phosphatase: use in topological studies with endoplasmic reticulum vesicles. Anal Biochem. 1992 Nov 1;206(2):328–333. doi: 10.1016/0003-2697(92)90374-g. [DOI] [PubMed] [Google Scholar]
- Rush J. S., Waechter C. J. Method for the determination of cellular levels of guanosine-5'-diphosphate-mannose based on a weak interaction with concanavalin A at low pH. Anal Biochem. 1995 Jan 20;224(2):494–501. doi: 10.1006/abio.1995.1078. [DOI] [PubMed] [Google Scholar]
- Sandhoff K., van Echten G. Ganglioside metabolism: enzymology, topology and regulation. Prog Brain Res. 1994;101:17–29. doi: 10.1016/s0079-6123(08)61937-8. [DOI] [PubMed] [Google Scholar]
- Schroit A. J., Zwaal R. F. Transbilayer movement of phospholipids in red cell and platelet membranes. Biochim Biophys Acta. 1991 Nov 13;1071(3):313–329. doi: 10.1016/0304-4157(91)90019-s. [DOI] [PubMed] [Google Scholar]
- Sharma C. B., Babczinski P., Lehle L., Tanner W. The role of dolicholmonophosphate in glycoprotein biosynthesis in Saccharomyces cerevisiae. Eur J Biochem. 1974 Jul 1;46(1):35–41. doi: 10.1111/j.1432-1033.1974.tb03594.x. [DOI] [PubMed] [Google Scholar]
- Stoll J., Robbins A. R., Krag S. S. Mutant of Chinese hamster ovary cells with altered mannose 6-phosphate receptor activity is unable to synthesize mannosylphosphoryldolichol. Proc Natl Acad Sci U S A. 1982 Apr;79(7):2296–2300. doi: 10.1073/pnas.79.7.2296. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Tanner W., Lehle L. Protein glycosylation in yeast. Biochim Biophys Acta. 1987 Apr 27;906(1):81–99. doi: 10.1016/0304-4157(87)90006-2. [DOI] [PubMed] [Google Scholar]
- Trotter P. J., Voelker D. R. Lipid transport processes in eukaryotic cells. Biochim Biophys Acta. 1994 Aug 4;1213(3):241–262. doi: 10.1016/0005-2760(94)00073-5. [DOI] [PubMed] [Google Scholar]
- Troy F. A., 2nd Polysialylation: from bacteria to brains. Glycobiology. 1992 Feb;2(1):5–23. doi: 10.1093/glycob/2.1.5. [DOI] [PubMed] [Google Scholar]
- Vidugiriene J., Menon A. K. The GPI anchor of cell-surface proteins is synthesized on the cytoplasmic face of the endoplasmic reticulum. J Cell Biol. 1994 Oct;127(2):333–341. doi: 10.1083/jcb.127.2.333. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Warren C. D., Jeanloz R. W. Chemical synthesis of dolichyl alpha-D-mannopyranosyl phosphate and citronellyl alpha-D-mannopyranosyl phosphate. Biochemistry. 1973 Dec 4;12(25):5038–5045. doi: 10.1021/bi00749a002. [DOI] [PubMed] [Google Scholar]
- Warren C. D., Jeanloz R. W. Chemical synthesis of dolichyl phosphate and dolichyl glycosyl phosphates and pyrophosphates or "dolichol intermediates". Methods Enzymol. 1978;50:122–137. doi: 10.1016/0076-6879(78)50010-4. [DOI] [PubMed] [Google Scholar]
- Weppner W. A., Neuhaus F. C. Biosynthesis of peptidoglycan. Definition of the microenvironment of undecaprenyl diphosphate-N-acetylmuramyl-(5-dimethylaminonaphthalene-1-sulfonyl) pentapeptide by fluorescence spectroscopy. J Biol Chem. 1978 Jan 25;253(2):472–478. [PubMed] [Google Scholar]
- Zachowski A., Devaux P. F. Transmembrane movements of lipids. Experientia. 1990 Jun 15;46(6):644–656. doi: 10.1007/BF01939703. [DOI] [PubMed] [Google Scholar]
- Zachowski A. Phospholipids in animal eukaryotic membranes: transverse asymmetry and movement. Biochem J. 1993 Aug 15;294(Pt 1):1–14. doi: 10.1042/bj2940001. [DOI] [PMC free article] [PubMed] [Google Scholar]