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. 1975 Feb;146(2):299–307. doi: 10.1042/bj1460299

Glycosylation of ribonuclease A catalysed by rabbit liver extracts.

Z Khalkhall, R D Marshall
PMCID: PMC1165306  PMID: 1156375

Abstract

Crude extracts of rabbit liver catalyse in vitro the transfer of N-acetylglucosamine from UDP-N-acetylglucosamine to bovine pancreatic ribonuclease A. The enzymic activity is contained in rough endoplasmic reticulum. It has an absolute requirement for a bivalent metal ion: Co-2+ greater than Mn-2+ greater than Ni-2+. Mg-2+ is ineffective. There is enzymic activity in the absence of detergent, but increased activity is observed in the presence of Triton X-100. The site of glycosylation of ribonuclease A is asparagine-34, and glycosylation occurs only at this point. These findings agree with the hypothesis that the sequence Asn-X-Thr(Ser) (where X may be one of a number of types of amino acid) is a necessary, but not sufficient, condition for N-acetylglucosaminylation of a protein-bound asparagine residue.

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Selected References

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  1. ATFIELD G. N., MORRIS C. J. Analytical separations by highvoltage paper electrophoresis. Amino acids in protein hydrolysates. Biochem J. 1961 Dec;81:606–614. doi: 10.1042/bj0810606. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Behrens N. H., Carminatti H., Staneloni R. J., Leloir L. F., Cantarella A. I. Formation of lipid-bound oligosaccharides containing mannose. Their role in glycoprotein synthesis. Proc Natl Acad Sci U S A. 1973 Dec;70(12):3390–3394. doi: 10.1073/pnas.70.12.3390. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Catley B. J., Moore S., Stein W. H. The carbohydrate moiety of bovine pancreatic deoxyribonuclease. J Biol Chem. 1969 Feb 10;244(3):933–936. [PubMed] [Google Scholar]
  4. DALLNER G. STUDIES ON THE STRUCTURAL AND ENZYMIC ORGANIZATION OF THE MEMBRANOUS ELEMENTS OF LIVER MICROSOMES. Acta Pathol Microbiol Scand Suppl. 1963:SUPPL166–SUPPL16694. [PubMed] [Google Scholar]
  5. Hunt L. T., Dayhoff M. O. The occurrence in proteins of the tripeptides Asn-X-Ser and Asn-X-Thr and of bound carbohydrate. Biochem Biophys Res Commun. 1970 May 22;39(4):757–765. doi: 10.1016/0006-291x(70)90270-6. [DOI] [PubMed] [Google Scholar]
  6. Jackson R. L., Hirs C. H. The primary structure of porcine pancreatic ribonuclease. I. The distribution and sites of carbohydrate attachment. J Biol Chem. 1970 Feb 10;245(3):624–636. [PubMed] [Google Scholar]
  7. KALNITSKY G., HUMMEL J. P., DIERKS C. [Some factors which affect the enzymatic digestion of ribonucleic acid]. J Biol Chem. 1959 Jun;234(6):1512–1516. [PubMed] [Google Scholar]
  8. Kono M., Yamashina I. Identification of asparagine as the site of glycosylation in glycoprotein biosynthesis. J Biochem. 1973 May;73(5):1089–1094. doi: 10.1093/oxfordjournals.jbchem.a130163. [DOI] [PubMed] [Google Scholar]
  9. Lawford G. R., Schachter H. Biosynthesis of glycoprotein by liver. The incorporation in vivo of 14C-glucosamine into protein-bound hexosamine and sialic acid of rat liver subcellular fractions. J Biol Chem. 1966 Nov 25;241(22):5408–5418. [PubMed] [Google Scholar]
  10. Leloir L. F. Two decades of research on the biosynthesis of saccharides. Science. 1971 Jun 25;172(3990):1299–1303. doi: 10.1126/science.172.3990.1299. [DOI] [PubMed] [Google Scholar]
  11. Letts P. J., Schachter H. Stimulation of rat liver microsomal UDP-N-acetylglucosamine: glycoprotein N-acetylglucosaminyltransferase by carboxymethylcysteine-ribonuclease A. Can J Biochem. 1973 Jan;51(1):101–105. doi: 10.1139/o73-012. [DOI] [PubMed] [Google Scholar]
  12. MAROUX S., ROVERY M., DESNUELLE P. [Purification of trypsinogen and bovine trypsin by chromatography on carboxymethyl cellulose]. Biochim Biophys Acta. 1962 Jan 1;56:202–204. doi: 10.1016/0006-3002(62)90556-5. [DOI] [PubMed] [Google Scholar]
  13. Marshall R. D. Glycoproteins. Annu Rev Biochem. 1972;41:673–702. doi: 10.1146/annurev.bi.41.070172.003325. [DOI] [PubMed] [Google Scholar]
  14. Marshall R. D. The nature and metabolism of the carbohydrate-peptide linkages of glycoproteins. Biochem Soc Symp. 1974;(40):17–26. [PubMed] [Google Scholar]
  15. Marshall R. D., Zamecnik P. C. Some physical properties of lysyl and arginyl-transfer RNA synthetases of Escherichia coli B. Biochim Biophys Acta. 1969 Jul 1;181(2):454–464. doi: 10.1016/0005-2795(69)90279-7. [DOI] [PubMed] [Google Scholar]
  16. Melchers F. The attachment site of carbohydrate in a mouse immunoglobulin light chain. Biochemistry. 1969 Mar;8(3):938–947. doi: 10.1021/bi00831a026. [DOI] [PubMed] [Google Scholar]
  17. Molnar J., Sy D. Attachment of glucosamine to protein at the ribosomal site of rat liver. Biochemistry. 1967 Jul;6(7):1941–1947. doi: 10.1021/bi00859a009. [DOI] [PubMed] [Google Scholar]
  18. Mookerjea S., Cole D. E., Chow A., Letts P. A role of cytidine 5'-diphosphocholine in the transfer of N-acetyl-glucosamine from UDP-N-acetylglucosamine into endogenous acceptor lipids and proteins in rat and hen liver microsomes. Can J Biochem. 1972 Oct;50(10):1094–1108. doi: 10.1139/o72-150. [DOI] [PubMed] [Google Scholar]
  19. Mookerjea S. Glycoprotein biosynthesis: stimulation of N-acetylglucosaminyltransferase activity by cytidine 5'-diphosphocholine. Can J Biochem. 1972 Oct;50(10):1082–1093. doi: 10.1139/o72-149. [DOI] [PubMed] [Google Scholar]
  20. PLUMMER T. H., Jr, HIRS C. H. ON THE STRUCTURE OF BOVINE PANCREATIC RIBONUCLEASE B. ISOLATION OF A GLYCOPEPTIDE. J Biol Chem. 1964 Aug;239:2530–2538. [PubMed] [Google Scholar]
  21. Redman C. M., Cherian M. G. The secretory pathways of rat serum glycoproteins and albumin. Localization of newly formed proteins within the endoplasmic reticulum. J Cell Biol. 1972 Feb;52(2):231–245. doi: 10.1083/jcb.52.2.231. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. 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]
  23. Robinson G. B. The role of polyribosomes in the biosynthesis of glycoprotein. Biochem J. 1969 Dec;115(5):1077–1078. doi: 10.1042/bj1151077. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. SMYTH D. G., STEIN W. H., MOORE S. The sequence of amino acid residues in bovine pancreatic ribonuclease: revisions and confirmations. J Biol Chem. 1963 Jan;238:227–234. [PubMed] [Google Scholar]
  25. TABORSKY G. Chromatography of ribonuclease on carboxymethyl cellulose columns. J Biol Chem. 1959 Oct;234:2652–2656. [PubMed] [Google Scholar]
  26. Tait G. H. Glycine decarboxylase in Rhodopseudomonas spheroides and in rat liver mitochondria. Biochem J. 1970 Aug;118(5):819–830. doi: 10.1042/bj1180819. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Tarentino A. L., Plummer T. H., Jr, Maley F. The release of intact oligosaccharides from specific glycoproteins by endo-beta-N-acetylglucosaminidase H. J Biol Chem. 1974 Feb 10;249(3):818–824. [PubMed] [Google Scholar]
  28. 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]

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