title: Conformational analysis of the Streptococcus pneumoniae hyaluronate lyase and characterization of its hyaluronan-specific carbohydrate-binding module.
abstract: For a subset of pathogenic microorganisms, including Streptococcus pneumoniae, the recognition and degradation of host hyaluronan contributes to bacterial spreading through the extracellular matrix and enhancing access to host cell surfaces. The hyaluronate lyase (Hyl) presented on the surface of S. pneumoniae performs this role. Using glycan microarray screening, affinity electrophoresis, and isothermal titration calorimetry we show that the N-terminal module of Hyl is a hyaluronan-specific carbohydrate-binding module (CBM) and the founding member of CBM family 70. The 1.2 Å resolution x-ray crystal structure of CBM70 revealed it to have a β-sandwich fold, similar to other CBMs. The electrostatic properties of the binding site, which was identified by site-directed mutagenesis, are distinct from other CBMs and complementary to its acidic ligand, hyaluronan. Dynamic light scattering and solution small angle x-ray scattering revealed the full-length Hyl protein to exist as a monomer/dimer mixture in solution. Through a detailed analysis of the small angle x-ray scattering data, we report the pseudoatomic solution structures of the monomer and dimer forms of the full-length multimodular Hyl.
authors.affiliation: Department of Biochemistry and Microbiology, University of Victoria, Victoria, British Columbia V8W 3P6, Canada.
authors.forename: Michael D L
authors.initials: MDL
authors.lastname: Suits
authors.affiliation: Department of Biochemistry and Microbiology, University of Victoria, Victoria, British Columbia V8W 3P6, Canada.
authors.forename: Benjamin
authors.initials: B
authors.lastname: Pluvinage
authors.affiliation: Department of Biochemistry and Microbiology, University of Victoria, Victoria, British Columbia V8W 3P6, Canada.
authors.forename: Adrienne
authors.initials: A
authors.lastname: Law
authors.affiliation: Glycosciences Laboratory, Imperial College London, Burlington Danes Building, Du Cane Road, London W12 0NN, United Kingdom, and.
authors.forename: Yan
authors.initials: Y
authors.lastname: Liu
authors.affiliation: Glycosciences Laboratory, Imperial College London, Burlington Danes Building, Du Cane Road, London W12 0NN, United Kingdom, and; REQUIMTE, Department of Chemistry, Faculdade de Ciências e Tecnologia, Universidade Nova de Lisboa, 2829-516 Caparica, Portugal.
authors.forename: Angelina S
authors.initials: AS
authors.lastname: Palma
authors.affiliation: Glycosciences Laboratory, Imperial College London, Burlington Danes Building, Du Cane Road, London W12 0NN, United Kingdom, and.
authors.forename: Wengang
authors.initials: W
authors.lastname: Chai
authors.affiliation: Glycosciences Laboratory, Imperial College London, Burlington Danes Building, Du Cane Road, London W12 0NN, United Kingdom, and.
authors.forename: Ten
authors.initials: T
authors.lastname: Feizi
authors.affiliation: Department of Biochemistry and Microbiology, University of Victoria, Victoria, British Columbia V8W 3P6, Canada,. Electronic address: boraston@uvic.ca.
authors.forename: Alisdair B
authors.initials: AB
authors.lastname: Boraston
mesh_terms: D001426:Bacterial Proteins / Q000737:chemistry* / Q000235:genetics; D018360:Crystallography, X-Ray; D016297:Mutagenesis, Site-Directed; D011133:Polysaccharide-Lyases / Q000737:chemistry* / Q000235:genetics; D055503:Protein Multimerization; D020836:Protein Structure, Quaternary; D017434:Protein Structure, Tertiary; D013296:Streptococcus pneumoniae / Q000201:enzymology* / Q000235:genetics
chemical_list: D001426:Bacterial Proteins; D011133:Polysaccharide-Lyases; C100002:hyaluronate lyase
keywords: Carbohydrate; Carbohydrate Processing; Carbohydrate-binding Protein; Crystallography; Glycobiology; Hyaluronan; Hyaluronate Lyase; Small Angle X-ray Scattering (SAXS); Streptococcus
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