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The Mechanism of Synthesis of a Mixed-Linkage (1right-arrow 3),(1right-arrow 4)beta -D-Glucan in Maize. Evidence for Multiple Sites of Glucosyl Transfer in the Synthase Complex1

Marcos S. Buckeridge, Claudia E. Vergara, and Nicholas C. Carpita*

Instituto de Botânica, Secão de Fisiologia e Bioquímica Plantas, Caixa Postal 4005, CEP-01061970, São Paulo, SP Brazil (M.S.B.); and Department of Botany and Plant Pathology, Purdue University, West Lafayette, Indiana 47907-1155 (C.E.V., N.C.C.)

We examined the mechanism of synthesis in vitro of (1right-arrow3),(1right-arrow4)beta -D-glucan (beta -glucan), a growth-specific cell wall polysaccharide found in grasses and cereals. beta -Glucan is composed primarily of cellotriosyl and cellotetraosyl units linked by single (1right-arrow3)beta -linkages. The ratio of cellotriosyl and cellotetraosyl units in the native polymer is strictly controlled at between 2 and 3 in all grasses, whereas the ratios of these units in beta -glucan formed in vitro vary from 1.5 with 5 µM UDP-glucose (Glc) to over 11 with 30 mM substrate. These results support a model in which three sites of glycosyl transfer occur within the synthase complex to produce the cellobiosyl-(1right-arrow3)-D-glucosyl units. We propose that failure to fill one of the sites results in the iterative addition of one or more cellobiosyl units to produce the longer cellodextrin units in the polymer. Variations in the UDP-Glc concentration in excised maize (Zea mays) coleoptiles did not result in wide variations in the ratios of cellotriosyl and cellotetraosyl units in beta -glucan synthesized in vivo, indicating that other factors control delivery of UDP-Glc to the synthase. In maize sucrose synthase is enriched in Golgi membranes and plasma membranes and may be involved in the control of substrate delivery to beta -glucan synthase and cellulose synthase.

1   This work was supported by contract DE-FG02-88ER13903 from the U.S. Department of Energy, Energy Biosciences (to N.C.C.) and a grant from Fundação de Amparo à Pesquisa do Estado de São Paulo (to M.S.B.). This is journal paper 16,002 of the Purdue Agricultural Experiment Station.
*   Corresponding author; e-mail carpita{at}btny.purdue.edu; fax 765-494-0363.

Plant Physiol. (1999) 120: 1105-1116
Copyright Clearance Center:   0032-0889/99/120//12
© 1999 American Society of Plant Physiologists




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