Skip to main content
NCBI home page
Biochemical Journal logoLink to Biochemical Journal
. 1988 Jul 15;253(2):577–580. doi: 10.1042/bj2530577

Purification of alpha 2,6-sialyltransferase from rat liver by dye chromatography.

U Sticher 1, H J Gross 1, R Brossmer 1
PMCID: PMC1149336  PMID: 3052425

Abstract

We describe a simple three-step purification for Gal-beta 1,4-GlcNAc-alpha 2,6-sialyltransferase (EC 2.4.99.1) from rat liver which uses chromatography on Cibacron Blue F3GA and f.p.l.c. It gives a highly purified (11,000-fold) enzyme in 19% yield, which is free of other sialyltransferases, CMP-NeuAc hydrolase, sialidases and proteinases.

Full text

PDF
577

Selected References

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

  1. Alhadeff J. A., Holzinger R. T. Purification of human liver glycoprotein sialyltransferase by affinity chromatography. J Biochem Biophys Methods. 1982 Aug;6(3):229–233. doi: 10.1016/0165-022x(82)90045-8. [DOI] [PubMed] [Google Scholar]
  2. Ashwell G., Morell A. G. The role of surface carbohydrates in the hepatic recognition and transport of circulating glycoproteins. Adv Enzymol Relat Areas Mol Biol. 1974;41(0):99–128. doi: 10.1002/9780470122860.ch3. [DOI] [PubMed] [Google Scholar]
  3. Dean P. D., Watson D. H. Protein purification using immobilised triazine dyes. J Chromatogr. 1979 Oct 1;165(3):301–319. doi: 10.1016/s0021-9673(00)88187-x. [DOI] [PubMed] [Google Scholar]
  4. Devi S. K., Moore R., Davids N., Findeis A. F., Berger R. L. Simulation of water movement across the cell membrane in a hypertonic medium. Comput Biol Med. 1970 Dec;1(2):141–153. doi: 10.1016/0010-4825(70)90006-5. [DOI] [PubMed] [Google Scholar]
  5. Gross H. J., Bünsch A., Paulson J. C., Brossmer R. Activation and transfer of novel synthetic 9-substituted sialic acids. Eur J Biochem. 1987 Nov 2;168(3):595–602. doi: 10.1111/j.1432-1033.1987.tb13458.x. [DOI] [PubMed] [Google Scholar]
  6. Joziasse D. H., Bergh M. L., ter Hart H. G., Koppen P. L., Hooghwinkel G. J., Van den Eijnden D. H. Purification and enzymatic characterization of CMP-sialic acid: beta-galactosyl1----3-N-acetylgalactosaminide alpha 2----3-sialyltransferase from human placenta. J Biol Chem. 1985 Apr 25;260(8):4941–4951. [PubMed] [Google Scholar]
  7. Kornfeld R., Kornfeld S. Comparative aspects of glycoprotein structure. Annu Rev Biochem. 1976;45:217–237. doi: 10.1146/annurev.bi.45.070176.001245. [DOI] [PubMed] [Google Scholar]
  8. Laemmli U. K. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970 Aug 15;227(5259):680–685. doi: 10.1038/227680a0. [DOI] [PubMed] [Google Scholar]
  9. Paulson J. C., Beranek W. E., Hill R. L. Purification of a sialyltransferase from bovine colostrum by affinity chromatography on CDP-agarose. J Biol Chem. 1977 Apr 10;252(7):2356–2362. [PubMed] [Google Scholar]
  10. Potier M., Mameli L., Bélisle M., Dallaire L., Melançon S. B. Fluorometric assay of neuraminidase with a sodium (4-methylumbelliferyl-alpha-D-N-acetylneuraminate) substrate. Anal Biochem. 1979 Apr 15;94(2):287–296. doi: 10.1016/0003-2697(79)90362-2. [DOI] [PubMed] [Google Scholar]
  11. Sadler J. E., Rearick J. I., Hill R. L. Purification to homogeneity and enzymatic characterization of an alpha-N-acetylgalactosaminide alpha 2 leads to 6 sialyltransferase from porcine submaxillary glands. J Biol Chem. 1979 Jul 10;254(13):5934–5941. [PubMed] [Google Scholar]
  12. Twining S. S. Fluorescein isothiocyanate-labeled casein assay for proteolytic enzymes. Anal Biochem. 1984 Nov 15;143(1):30–34. doi: 10.1016/0003-2697(84)90553-0. [DOI] [PubMed] [Google Scholar]
  13. Weinstein J., de Souza-e-Silva U., Paulson J. C. Purification of a Gal beta 1 to 4GlcNAc alpha 2 to 6 sialyltransferase and a Gal beta 1 to 3(4)GlcNAc alpha 2 to 3 sialyltransferase to homogeneity from rat liver. J Biol Chem. 1982 Nov 25;257(22):13835–13844. [PubMed] [Google Scholar]
  14. Weinstein J., de Souza-e-Silva U., Paulson J. C. Sialylation of glycoprotein oligosaccharides N-linked to asparagine. Enzymatic characterization of a Gal beta 1 to 3(4)GlcNAc alpha 2 to 3 sialyltransferase and a Gal beta 1 to 4GlcNAc alpha 2 to 6 sialyltransferase from rat liver. J Biol Chem. 1982 Nov 25;257(22):13845–13853. [PubMed] [Google Scholar]
  15. Williams H. R., Lin T. Y. Methyl- 14 C-glycinated hemoglobin as a substrate for proteases. Biochim Biophys Acta. 1971 Dec 15;250(3):603–607. doi: 10.1016/0005-2744(71)90265-8. [DOI] [PubMed] [Google Scholar]

Articles from Biochemical Journal are provided here courtesy of The Biochemical Society

RESOURCES