Structure-function relationships for Schizophyllum commune trehalose phosphorylase and their implications for the catalytic mechanism of family GT-4 glycosyltransferases.

The Biochemical Journal
Christiane GoedlBernd Nidetzky

Abstract

The cDNA encoding trehalose phosphorylase, a family GT-4 glycosyltransferase from the fungus Schizophyllum commune, was isolated and expressed in Escherichia coli to yield functional recombinant protein in its full length of 737 amino acids. Unlike the natural phosphorylase that was previously obtained as a truncated 61 kDa monomer containing one tightly bound Mg2+, the intact enzyme produced in E. coli is a dimer and not associated with metal ions [Eis, Watkins, Prohaska and Nidetzky (2001) Biochem. J. 356, 757-767]. MS analysis of the slow spontaneous conversion of the full-length enzyme into a 61 kDa fragment that is fully active revealed that critical elements of catalysis and specificity of trehalose phosphorylase reside entirely in the C-terminal protein part. Intact and truncated phosphorylases thus show identical inhibition constants for the transition state analogue orthovanadate and alpha,alpha-trehalose (K(i) approximately 1 microM). Structure-based sequence comparison with retaining glycosyltransferases of fold family GT-B reveals a putative active centre of trehalose phosphorylase, and results of site-directed mutagenesis confirm the predicted crucial role of Asp379, His403, Arg507 and Lys512 in catalysis and also ...Continue Reading

References

Jan 1, 1983·Molecular and Cellular Biochemistry·M Mojena, J E Felíu
Sep 15, 1996·European Journal of Biochemistry·N AsanoK Matsui
Oct 9, 1998·Applied Microbiology and Biotechnology·K SaitoS Horinouchi
Sep 26, 2000·The Journal of Biological Chemistry·P L AbdianR A Geremia
Sep 10, 2002·Journal of Molecular Biology·S GeremiaL N Johnson
Nov 5, 2002·Current Opinion in Chemical Biology·Andrea VasellaMatthias Böhm
Apr 15, 2003·Journal of Molecular Biology·Pedro M CoutinhoBernard Henrissat
Jul 26, 2003·Protein Expression and Purification·Sang-Eun HanMyung-Ok Byun
Dec 11, 2003·The Journal of Biological Chemistry·Alejandra YepJack Preiss
Mar 23, 2004·Biochemical and Biophysical Research Communications·Alejandra YepJack Preiss
Oct 19, 2004·Chemical Communications : Chem Comm·Luke L Lairson, Stephen G Withers
Mar 2, 2005·Biochemical and Biophysical Research Communications·Soo-In RyuSoo-Bok Lee
Jun 14, 2005·Nature Structural & Molecular Biology·James FlintGideon J Davies
Jul 23, 2005·Glycobiology·Christelle BretonAnne Imberty
Aug 6, 2005·Journal of Molecular Biology·Laurent LarivièreSolange Moréra
Nov 3, 2005·Current Opinion in Structural Biology·Gideon J DaviesBernard Henrissat

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Citations

Aug 2, 2011·Applied and Environmental Microbiology·Jef Van der BorghtWim Soetaert
Aug 9, 2011·Nature Chemical Biology·Seung Seo LeeBenjamin G Davis
Dec 15, 2015·Biotechnology Advances·Katharina SchmölzerBernd Nidetzky
Feb 19, 2008·Carbohydrate Research·Lothar BreckerBernd Nidetzky
Feb 10, 2019·Applied and Environmental Microbiology·Luuk MestromPeter-Leon Hagedoorn
Dec 12, 2017·Pure and Applied Chemistry. Chimie Pure Et Appliquée·Mara K O'NeillBenjamin M Swarts
Jun 18, 2015·International Journal of Molecular Sciences·Maarten WalmaghTom Desmet
Dec 15, 2006·Journal of the American Chemical Society·Ana Monegal, Antoni Planas

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