Single-molecular pair unbinding studies of Mannuronan C-5 epimerase AlgE4 and its polymer substrate

Biomacromolecules
Marit SletmoenB T Stokke

Abstract

Alginate biosynthesis involves C-5-mannuronan epimerases catalyzing the conversion of beta-D-mannuronic acid to alpha-L-guluronic acid at the polymer level. Mannuronan epimerases are modular enzymes where the various modules yield specific sequential patterns of the converted residues in their polymer products. Here, the interaction between the AlgE4 epimerase and mannuronan is determined by dynamic force spectroscopy. The specific unbinding between molecular pairs of mannuronan and AlgE4 as well as its two modules, A and R, respectively, was studied as a function of force loading rate. The mean protein-mannuronan unbinding forces were determined to be in the range 73-144 pN, depending on the protein, at a loading rate of 0.6 nN/s, and increased with increasing loading rate. The position of the activation barrier was determined to be 0.23 +/- 0.04 nm for the AlgE4 and 0.10 +/- 0.02 nm for its A-module. The lack of interaction observed between the R-module and mannuronan suggest that the A-module contains the binding site for the polymer substrate. The ratio between the epimerase-mannuronan dissociation rate and the catalytic rate for epimerization of single hexose residues suggests a processive mode of action of the AlgE4 epime...Continue Reading

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Citations

Feb 1, 2012·Journal of Molecular Recognition : JMR·Dionne C G KleinBjørn T Stokke
Oct 4, 2006·Micron : the International Research and Review Journal for Microscopy·Chih-Kung LeeShiming Lin
Mar 13, 2012·International Journal of Molecular Sciences·Katarzyna Maria Psonka-AntonczykMikael Lindgren
Jan 13, 2006·The Journal of Biological Chemistry·Finn L AachmannReinhard Wimmer
Jan 9, 2014·The Journal of Biological Chemistry·Francis WolframP Lynne Howell
Jun 9, 2020·Frontiers in Molecular Biosciences·Byeongseon YangMichael A Nash
Aug 19, 2017·Scientific Reports·K K SandJ J DeYoreo
Jun 6, 2009·Langmuir : the ACS Journal of Surfaces and Colloids·Makoto TakemasaBjørn T Stokke
Apr 19, 2007·Langmuir : the ACS Journal of Surfaces and Colloids·Chad RayBoris B Akhremitchev

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