Plant Physiol.
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Plant Physiology 70:12-20 (1982)
© 1982 American Society of Plant Biologists

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Articles

Characterization of the Oligosaccharides from Lipid-Linked Oligosaccharides of Mung Bean Seedlings 1

Hidetaka Hori and Alan D. Elbein

Department of Biochemistry, The University of Texas Health Science Center, San Antonio, Texas 78284

Lipid-linked oligosaccharides were synthesized with the particulate enzyme preparation from mung bean (Phaseolus aureus) seedlings in the presence of GDP-[14C] mannose. The oligosaccharides were released from the lipids by mild acid hydrolysis and purified by several passages on Biogel P-4 columns. Five different oligosaccharides were purified in this way. Based on their relative elution constants (Kd) compared to a variety of standard oligosaccharides, they were sized as (mannose-acetylglucosamine) Man7GlcNAc2, Man5GlcNAc2, Man3GlcNAc2, Man2GlcNAc2, and ManGlcNAc2. These oligosaccharides were treated with endoglucosaminidase H and {alpha}- and {beta}-mannosidase, and the products were examined on Biogel P-4 columns. They also were subjected to a number of chemical treatments including analysis of the reducing sugar by NaB3H4 reduction, methylation analysis, and in some cases acetolysis. From these data, the likely structures of these oligosaccharides are as follows: E, Man{beta}-GlcNAc-GlcNAc; D, Man{alpha}1->3Man{beta}-GlcNAc-GlcNAc; C, Man{alpha}1->2Man{alpha}1->3Man{beta}-GlcNAc-GlcNAc; B, Man{alpha}1->2Man{alpha}1->2Man{alpha}1-> 3(Man{alpha}1->6)Man{beta}-GlcNAc-GlcNAc; and A, Man{alpha}1->2Man{alpha}1-> 2Man{alpha}1->3(Man{alpha}1-> [Man{alpha}1->6]Man{alpha}1->6) Man{beta}-GlcNAc-GlcNAc. The synthesis of the Man7GlcNAc2 was greatly diminished when tunicamycin (10 µg/ml) was added to the incubation mixtures.

1 Supported by grants from the National Institutes of Health (AM 21800) and the Robert A. Welch Foundation.






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Copyright © 1982 by the American Society of Plant Biologists