Plant Physiol. Drug Metab Dispos
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Plant Physiology 81:913-918 (1986)
© 1986 American Society of Plant Biologists

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Gel-Electrophoretic Separation, Detection, and Characterization of Plant and Bacterial UDP-Glucose Glucosyltransferases

Michael P. Thelen1 and Deborah P. Delmer

ARCO Plant Cell Research Institute, 6560 Trinity Court, Dublin, California 94568

We have developed procedures for detection and characterization of UDP-glucose: glucosyltransferases following electrophoretic separation in nondenaturing polyacrylamide gels. Using digitonin-solubilized membrane protein preparations from a variety of plants and two cellulose-producing bacteria, activity can be demonstrated for several UDP-glucose:beta-glucan synthases with an in situ assay following gel electrophoresis. These enzymes can be characterized within the gels with respect to effector requirements and products produced, and several advantages of this assay over solution assays are demonstrated. For example, the clear dependence of plant UDP-glucose:(1->3)-beta-glucan synthase on both Ca2+ and a beta-linked glucoside is shown; bacterial cellulose synthases show direct stimulation within the gel by guanyl oligonucleotide, and the Acetobacter xylinum enzyme appears more stable in the gel assay than in solution assay.


1 Present address: Biochemistry Department, Tennis Court Road, University of Cambridge, Cambridge, CB2 1QW, England.




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Copyright © 1986 by the American Society of Plant Biologists