Visualization of the activity of xyloglucan endotransglycosylase (XET) isoenzymes after gel electrophoresis

P P M Iannetta, S C Fry

Research output: Contribution to journalArticlepeer-review

Abstract

A protocol is described which allows the detection of transglycosylase isoenzyme activity, Crude extracts of cauliflower (Brassica oleracea var. botrytis) fiorets were treated with 1 M sodium chloride, filtered through cellulose acetate, dialysed, concentrated and subjected to isoelectric focusing under non-denaturing conditions in a polyacrylamide gel. After electrophoresis, the gel was overlaid with a test paper impregnated with a mixture of high-M-r xyloglucan and a sulphorhodamine-conjugated oligosaccharide of xyloglucan. Transglycosylation caused the production of sulphorhodamine-labelled polysaccharide, which remained hound to the test paper after prolonged washing in water, revealing eight fluorescent hands corresponding to XET isoenzymes with pi values of 8.8, 8.6, 7.8, 7.6, 7.5, 7.1, 6,7 and 6.6. Copyright (C) 1999 John Wiley & Sons, Ltd.

Original languageEnglish
Pages (from-to)238-240
Number of pages3
JournalPhytochemical analysis
Volume10
Issue number5
Publication statusPublished - 1999

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