Immunocytochemical localization of α2,3(N)-sialyltransferase (ST3Gal III) in cell lines and rat kidney tissue sections:: evidence for Golgi and post-Golgi localization

被引:23
作者
Burger, PC
Lötscher, M
Streiff, M
Kleene, R
Kaissling, B
Berger, EG
机构
[1] Univ Zurich, Inst Physiol, CH-8057 Zurich, Switzerland
[2] Univ Zurich, Inst Anat, CH-8057 Zurich, Switzerland
[3] Novartis Pharma AG, CH-4002 Basel, Switzerland
关键词
alpha 2,3(N)-sialyltransferase; beta-galactosidase; Golgi apparatus; kidney tubules; Jurkat cells;
D O I
10.1093/glycob/8.3.245
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
Sialylation is a biosynthetic process occurring in the trans compartments of the Golgi apparatus. Corresponding evidence is based on localization and biochemical studies of alpha 2,6(N)-sialyltransferase (ST6Gal I) as previously reported. Here we describe generation and characterization of polyclonal antibodies to recombinant rat alpha 2,3(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 beta 1,4-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 in perfusion-fixed rat kidney sections. We found strong staining of the Golgi apparatus of tubular epithelia and a brush-border-associated staining which colocalized with cytochemical staining of the H(+)ATPase. This subcellular localization was not observed for ST6Gal I which localized to the Golgi apparatus. These data show colocalization in the Golgi apparatus and different post-Golgi distributions of the two sialyltransferases.
引用
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页码:245 / 257
页数:13
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