Bacterial polysaccharide lyase family 33: Specificity from an evolutionarily conserved binding tunnel
Abstract
Acidic glycans are essential for the biology of multicellular eukaryotes. To utilize them, microbial life including symbionts and pathogens has evolved polysaccharide lyases (PL) that cleave their 1,4 glycosidic linkages via a β-elimination mechanism. PL family 33 (PL33) enzymes have the unusual ability to target a diverse range of glycosaminoglycans (GAGs), as well as the bacterial polymer, gellan gum. In order to gain more detailed insight into PL33 activities we recombinantly expressed 10 PL33 members derived from all major environments and further elucidated the detailed biochemical and biophysical properties of five, showing that their substrate specificity is conferred by variations in tunnel length and topography. The key amino acids involved in catalysis and substrate interactions were identified, and employing a combination of complementary biochemical, structural, and modeling approaches, we show that the tunnel topography is induced by substrate binding to the glycan. Structural and bioinformatic analyses revealed that these features are conserved across several lyase families as well as in mammalian GAG epimerases.
Article Details
Journal Info
Proceedings of the National Academy of Sciences
National Academy of Sciences
Authors (19)
Mélanie Loiodice
Université Grenoble Alpes, CNRS, Centre de Recherche sur les Macromolécules Végétales
Elodie Drula
Architecture et Fonction des Macromolécules Biologiques, Aix-Marseille Univ, CNRS, Institut National de Recherche pour l'Agriculture, l'alimentation et l'Environnement
Zak McIver
Svetlana Antonyuk
Department of Biochemistry, Cell and Systems Biology, Institute of Systems, Molecular and Integrative Biology, University of Liverpool
Arnaud Basle
Marcelo Lima
School of Life Sciences, Keele University
Edwin A. Yates
Department of Biochemistry, Cell and Systems Biology, Institute of Systems, Molecular and Integrative Biology, University of Liverpool
Dominic P. Byrne
Department of Biochemistry, Cell and Systems Biology, Institute of Systems, Molecular and Integrative Biology, University of Liverpool
Jamie Coughlan
Department of Biochemistry, Cell and Systems Biology, Institute of Systems, Molecular and Integrative Biology, University of Liverpool
Andrew Leech
Department of Biology, Technology Facility, University of York
Shahram Mesdaghi
Department of Biochemistry, Cell and Systems Biology, Institute of Systems, Molecular and Integrative Biology, University of Liverpool
Daniel J. Rigden
Department of Biochemistry, Cell and Systems Biology, Institute of Systems, Molecular and Integrative Biology, University of Liverpool
Sophie Drouillard
Université Grenoble Alpes, CNRS, Centre de Recherche sur les Macromolécules Végétales
William Helbert
Université Grenoble Alpes, CNRS, Centre de Recherche sur les Macromolécules Végétales
Bernard Henrissat
Nicolas Terrapon
Gareth S. A. Wright
School of Life Sciences, University of Essex
Marie Couturier
Université Grenoble Alpes, CNRS, Centre de Recherche sur les Macromolécules Végétales
Alan Cartmell