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Journal of Cell Science, Vol 96, 705-710, Copyright © 1990 by Company of Biologists
Submitted on February 15, 1990
Accepted on April 17, 1990
1 Biology Department, McGill University, 1205 Avenue Dr Penfield, Montreal, Quebec, Canada H3A 1B1
2 McGill Cancer Centre, McGill University, 3665 Drummond Street, Montreal, Quebec, Canada H3G 1Y6
Author for correspondence at: Genetic Engineering of Proteins, Room 2129, National Research Council, 100 Sussex Drive, Ottawa, Ontario, Canada K1A 0R6
Rate-zonal centrifugation of pea (Pisum sativum var. Alaska) stem microsomal membranes on a linear Renografln gradient separated Golgi secretory vesicles from dictyosomes. Secretory vesicles possessed high levels of xyloglucan fucosyl transferase activity, which effects the final decoration of stem xyloglucan side-chains, but lacked substantial xyloglucan xylosyl transferase activity, which is required for the synthesis of the xyloglucan backbone. In contrast, total dictyosomal membranes possessed both fucosyl and xylosyl transferase activities. Isopycnic centrifugation of a homogenized dictyosome-enriched membrane preparation on a shallower Renografln gradient indicated that lighter dictyosomal membranes possessed xylosyl transferase but relatively little fucosyl transferase activity. The bulk of the dictyosomal membranes formed a denser peak in which xylosyl and fucosyl transferase activities codistributed. Thus a differential localization of function in the Golgi apparatus during biosynthesis of xyloglucan is indicated. A tentative mechanism is suggested in which the elaboration of the glucose-xylose backbone is initiated in lighter dictyosomal membranes, backbone synthesis is concluded and fucosylation begun in denser dictyosomal membranes, and fucosylation completed in Golgi secretory vesicles during transport of xyloglucan to the cell wall.
Key words: cell wall synthesis, Golgi, fucosyl transferase, xylosyl transferase, pea, xyloglucan, dictyosomes, secretory vesicles
Submitted on February 15, 1990
Accepted on April 17, 1990
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