Immunocytochemical localization of alpha2,3(N)-sialyltransferase (ST3Gal III) in cell lines and rat kidney tissue sections: evidence for golgi and post-golgi localization

Glycobiology
P C BurgerE G Berger

Abstract

Sialylation is a biosynthetic process occurring in the trans compartments of the Golgi apparatus. Corresponding evidence is based on localization and biochemical studies of alpha2, 6(N)-sialyltransferase (ST6Gal I) as previously reported. Here we describe generation and characterization of polyclonal antibodies to recombinant rat alpha2,3(N)-sialyltransferase (ST3Gal III) expressed as a soluble enzyme in Sf9 cells or as a beta-galactosidase-human-ST3Gal III fusion-protein from E.coli , respectively. These antibodies were used to localize ST3Gal III by immunofluorescence in various cell lines and rat kidney tissue sections. In transiently transfected COS cells the antibodies directed to soluble sialyltransferase or the sialyltransferase portion of the fusion-protein only recognized the recombinant antigen retained in the endoplasmic reticulum. However, an antibody fraction crossreactive with beta-galactosidase recognized natively expressed ST3Gal III which was found to be colocalized with beta1, 4-galactosyltransferase in the Golgi apparatus of several cultured cell lines. Antibodies affinity purified on the beta-galactosidase-ST3Gal III fusion-protein column derived from both antisera have then been used to localize the enzyme ...Continue Reading

Citations

Mar 12, 2002·Glycobiology·Eric G Berger
Apr 24, 2002·Analytical Biochemistry·Anthony H MerryPauline M Rudd
Jul 19, 2000·Proceedings of the National Academy of Sciences of the United States of America·S Schnyder-CandrianE G Berger
Mar 12, 2009·Journal of Cellular Physiology·Antti RivinojaSakari Kellokumpu
Sep 30, 2016·Medicinal Research Reviews·Rémi Szabo, Danielle Skropeta
Mar 22, 2001·The Journal of Histochemistry and Cytochemistry : Official Journal of the Histochemistry Society·F Ulloa, F X Real
Mar 30, 2001·The Journal of Biological Chemistry·P LegaigneurC Ronin
Sep 13, 2011·American Journal of Human Genetics·Hao HuAndreas W Kuss

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