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. 1975 May;148(2):245–251. doi: 10.1042/bj1480245

The formation of mono-N-acetylhexosamine derivatives of dolichol diphosphate by pig liver microsomal fractions.

G Palamarczyk, F W Hemming
PMCID: PMC1165532  PMID: 239708

Abstract

Incubation of pig liver microsomal preparations with UDP-N[U-14C]acetylglucosamine yields a 14C-labelled lipid. The requirement for Mn2+, the pH optimum, time-dependence and the reversibility by UMP of the transferase are reported. Evidence is presented in favour of the lipid being a mixture of dolichol diphosphate N-[14C]acetylglucosamine and dolichol diphosphate N-[14C]acetylmannosamine. Available data suggest that the epimerization takes place while the hexosamine is bound in this lipid-soluble form. The N-acetylmannosamine appeared not be be released into the medium. The subfractionation of the microsomal fraction to separate transferase activity from membrane-bound beta-N-acetylglucosaminidase activity is also reported.

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Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Barr R. M., Hemming F. W. Polyprenol phosphate as an acceptor of mannose from guanosine diphosphate mannose in Aspergillus niger. Biochem J. 1972 Mar;126(5):1203–1208. doi: 10.1042/bj1261203. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Behrens N. H., Parodi A. J., Leloir L. F., Krisman C. R. The role of dolichol monophosphate in sugar transfer. Arch Biochem Biophys. 1971 Apr;143(2):375–383. doi: 10.1016/0003-9861(71)90224-4. [DOI] [PubMed] [Google Scholar]
  3. Ehrenreich J. H., Bergeron J. J., Siekevitz P., Palade G. E. Golgi fractions prepared from rat liver homogenates. I. Isolation procedure and morphological characterization. J Cell Biol. 1973 Oct;59(1):45–72. doi: 10.1083/jcb.59.1.45. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Evans P. J., Hemming F. W. The unambiguous characterization of dolichol phosphate mannose as a product of mannosyl transferase in pig liver endoplasmic reticulum. FEBS Lett. 1973 May 1;31(3):335–338. doi: 10.1016/0014-5793(73)80135-8. [DOI] [PubMed] [Google Scholar]
  5. FOLCH J., LEES M., SLOANE STANLEY G. H. A simple method for the isolation and purification of total lipides from animal tissues. J Biol Chem. 1957 May;226(1):497–509. [PubMed] [Google Scholar]
  6. Ghalambor M. A., Warren C. D., Jeanloz R. W. Biosynthesis of a P1-2-acetamido-2-deoxy-D-glucosyl P2-polyisoprenyl pyrophosphate by calf pancreas microsomes. Biochem Biophys Res Commun. 1974 Jan 23;56(2):407–414. doi: 10.1016/0006-291x(74)90857-2. [DOI] [PubMed] [Google Scholar]
  7. LOWRY O. H., ROSEBROUGH N. J., FARR A. L., RANDALL R. J. Protein measurement with the Folin phenol reagent. J Biol Chem. 1951 Nov;193(1):265–275. [PubMed] [Google Scholar]
  8. Leloir L. F., Staneloni R. J., Carminatti H., Behrens N. H. The biosynthesis of a N,N'-diacetylchitobiose containing lipid by liver microsomes. A probable dolichol pyrophosphate derivative. Biochem Biophys Res Commun. 1973 Jun 19;52(4):1285–1292. doi: 10.1016/0006-291x(73)90640-2. [DOI] [PubMed] [Google Scholar]
  9. Levy J. A., Carminatti H., Cantarella A. I., Behrens N. H., Leloir L. F., Tábora E. Mannose transfer to lipid linked di-N-acetylchitobiose. Biochem Biophys Res Commun. 1974 Sep 9;60(1):118–125. doi: 10.1016/0006-291x(74)90180-6. [DOI] [PubMed] [Google Scholar]
  10. Molnar J., Chao H., Ikehara Y. Phosphoryl-N-acetylglucosamine transfer to a lipid acceptor of liver microsomal preparations. Biochim Biophys Acta. 1971 Sep 1;239(3):401–410. doi: 10.1016/0005-2760(71)90033-6. [DOI] [PubMed] [Google Scholar]
  11. Richards J. B., Hemming F. W. The transfer of mannose from guanosine diphosphate mannose to dolichol phosphate and protein by pig liver endoplasmic reticulum. Biochem J. 1972 Nov;130(1):77–93. doi: 10.1042/bj1300077. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Salo W. L., Fletcher H. G., Jr Studies onthe mechanism of action of uridine diphosphate N-acetylglucosamine 2-epimerase. Biochemistry. 1970 Feb 17;9(4):882–885. doi: 10.1021/bi00806a024. [DOI] [PubMed] [Google Scholar]
  13. Tetas M., Chao H., Molnar J. Incorporation of carbohydrates into endogenous acceptors of liver microsomal fractions. Arch Biochem Biophys. 1970 May;138(1):135–146. doi: 10.1016/0003-9861(70)90292-4. [DOI] [PubMed] [Google Scholar]
  14. Touster O., Aronson N. N., Jr, Dulaney J. T., Hendrickson H. Isolation of rat liver plasma membranes. Use of nucleotide pyrophosphatase and phosphodiesterase I as marker enzymes. J Cell Biol. 1970 Dec;47(3):604–618. doi: 10.1083/jcb.47.3.604. [DOI] [PMC free article] [PubMed] [Google Scholar]

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