Characterization of UDP-glucose:protein transglucosylase genes from potato

Characterization of UDP-glucose:protein transglucosylase genes from potato Many plant autocatalytic glycosyltransferases are implicated in plant polysaccharide biosynthesis. Cloning of cDNAs encoding potato (Solanum tuberosum L.) UDP-Glc:protein transglucosylase (UPTG, EC 2.4.1.112) and expression of the cDNA clone E11 in Escherichia coli have been previously reported. Here, we studied the functional expression of a second cDNA of the enzyme (E2 clone). Northern blots analysis, with specific cDNA probes for the two UPTG isoforms, showed a differential expression pattern of mRNA levels in different potato tissues. Moreover, both UPTG recombinant enzymes showed different kinetic parameters. The recombinant protein encoded by E2 clone has an apparent Imax for UDP-Xyl and UDP-Gal, significantly higher than for UDP-Glc. The K m values for UDP-Glc were 0.45–0.71 μM and the values for UDP-Xyl and UDP-Gal were slightly higher than that of the UDP-Glc (1.2–2.71 μM) for both UPTG recombinant enzymes. The present study revealed further evidence for the proposed role of UPTG in the synthesis of cell wall polysaccharide. It was found a correlation between UPTG transcript levels and the growing state of the tissues in which there was an active synthesis of cell wall components. Southern blot analysis indicates that at least three genes encoding UPTG are present in potato genome. Phylogenetic analysis of both UPTG recombinant proteins showed that they are members of the RGP subfamilies from dicots. http://www.deepdyve.com/assets/images/DeepDyve-Logo-lg.png Plant Molecular Biology Springer Journals

Characterization of UDP-glucose:protein transglucosylase genes from potato

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Publisher
Kluwer Academic Publishers
Copyright
Copyright © 2003 by Kluwer Academic Publishers
Subject
Life Sciences; Biochemistry, general; Plant Sciences; Plant Pathology
ISSN
0167-4412
eISSN
1573-5028
D.O.I.
10.1023/A:1025061324856
Publisher site
See Article on Publisher Site

Abstract

Many plant autocatalytic glycosyltransferases are implicated in plant polysaccharide biosynthesis. Cloning of cDNAs encoding potato (Solanum tuberosum L.) UDP-Glc:protein transglucosylase (UPTG, EC 2.4.1.112) and expression of the cDNA clone E11 in Escherichia coli have been previously reported. Here, we studied the functional expression of a second cDNA of the enzyme (E2 clone). Northern blots analysis, with specific cDNA probes for the two UPTG isoforms, showed a differential expression pattern of mRNA levels in different potato tissues. Moreover, both UPTG recombinant enzymes showed different kinetic parameters. The recombinant protein encoded by E2 clone has an apparent Imax for UDP-Xyl and UDP-Gal, significantly higher than for UDP-Glc. The K m values for UDP-Glc were 0.45–0.71 μM and the values for UDP-Xyl and UDP-Gal were slightly higher than that of the UDP-Glc (1.2–2.71 μM) for both UPTG recombinant enzymes. The present study revealed further evidence for the proposed role of UPTG in the synthesis of cell wall polysaccharide. It was found a correlation between UPTG transcript levels and the growing state of the tissues in which there was an active synthesis of cell wall components. Southern blot analysis indicates that at least three genes encoding UPTG are present in potato genome. Phylogenetic analysis of both UPTG recombinant proteins showed that they are members of the RGP subfamilies from dicots.

Journal

Plant Molecular BiologySpringer Journals

Published: Oct 7, 2004

References

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