PMID: 701363Sep 1, 1978Paper

Spatial orientation of glycoproteins in membranes of rat liver rough microsomes. I. Localization of lectin-binding sites in microsomal membranes

The Journal of Cell Biology
E Rodriguez BoulanD D Sabatini

Abstract

Carbohydrate-containing structures in rat liver rough microsomes (RM) were localized and characterized using iodinated lectins of defined specificity. Binding of [125I]Con A increased six- to sevenfold in the presence of low DOC (0.04--0.05%) which opens the vesicles and allows the penetration of the lectins. On the other hand, binding of [125I]WGA and [125I]RCA increased only slightly when the microsomal vesicles were opened by DOC. Sites available in the intact microsomal fraction had an affinity for [125I]Con A 14 times higher than sites for lectin binding which were exposed by the detergent treatment. Lectin-binding sites in RM were also localized electron microscopically with lectins covalently bound to biotin, which, in turn, were visualized after their reaction with ferritin-avidin (F-Av) markers. Using this method, it was demonstrated that in untreated RM samples, binding sites for lectins are not present on the cytoplasmic face of the microsomal vesicles, even after removal of ribosomes by treatment with high salt buffer and puromycin, but are located on smooth membranes which contaminate the rough microsomal fraction. Combining this technique with procedures which render the interior of the microsomal vesicles accessi...Continue Reading

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Citations

Mar 1, 1982·Journal of Ultrastructure Research·M Dauwalder, W G Whaley
Feb 1, 1988·Proceedings of the National Academy of Sciences of the United States of America·C Abeijon, C B Hirschberg
Mar 1, 1989·Proceedings of the National Academy of Sciences of the United States of America·M E Milla, C B Hirschberg
Nov 1, 1985·The Journal of Cell Biology·R BrandsJ E Rothman
Dec 1, 1982·The Journal of Cell Biology·L GeraceC Kondor-Koch
Jan 1, 1984·The Journal of Cell Biology·M R Torrisi, P Pinto da Silva
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Jun 1, 1983·Journal of Cellular Physiology·A D ElbeinK Vosbeck
May 15, 1988·Analytical Biochemistry·M Wilchek, E A Bayer
Feb 1, 1982·Journal of Dairy Science·J C Mercier, P Gaye
Dec 24, 1982·Philosophical Transactions of the Royal Society of London. Series B, Biological Sciences·A M Tartakoff
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Jan 20, 1987·Biochimica Et Biophysica Acta·T Brody, J W Suttie
Oct 5, 1987·Biochimica Et Biophysica Acta·J Roth
Apr 1, 1983·Molecular and Cellular Biology·A Ben-Ze'ev, R Abulafia

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