More than ten b-glycosyltransferases are now recognized that have limited similarity to the amino acid sequence of cellulose synthase from Acetobacter xylinum. Using hydrophobic cluster analysis (HCA), we recently identified two domains and putative catalytic residues in the processive b-glycosyltransferases. In this study, we have found expressed sequence tags (ESTs) from higher plants (Arabidopsis thaliana, Brassica campestris, and Oryza sativa) that exhibit a limited sequence similarity to the A. xylinum cellulose synthase. These ESTs contain some of the conserved residues identified in the processive b-glycosyltransferases. Complete sequencing of an EST clone (T88271) from A.thaliana led to the identification of all the conserved residues in the derived truncated polypeptide which appears to be part of a putative cellulose synthase. Sequence comparison of proteins with known function and several unidentified proteins have the ‘D, D, D35Q(R,Q)XRW’motif which is considered a strong predictor for â-glycosyltransferases that includes, among other proteins, cellulose and chitin synthases. The first two conserved aspartic acid residues in this motif were analysed by site-directed mutagenesis, and their replacement by another amino acid led to a loss of cellulose synthase activity in A.xylinum, suggesting that they are essential for enzyme activity. A correlation between the second residue (R or Q) in the Q(R,Q)XRW sequence and the synthesis of a long glucan chain (polysaccharide) or a short glucan chain (oligosaccharide) suggests that this residue may be involved in the degree of processivity.
KEYWORDS: b-Glycosyltransferases, Acetobacter xylinum, cellulose synthase, higher plants
For Online Viewing (2 Mb)
For Printing (3 Mb)
Up to the 1997 Publications Page
Up to Malcolm Brown's Lab Page
Last modified March 18, 2008
This document is maintained by R. Malcolm Brown, Jr.