Structural basis for the recognition and cleavage of polysialic acid by the bacteriophage K1F tailspike protein EndoNF.

Abstract

An alpha-2,8-linked polysialic acid (polySia) capsule confers immune tolerance to neuroinvasive, pathogenic prokaryotes such as Escherichia coli K1 and Neisseria meningitidis and supports host infection by means of molecular mimicry. Bacteriophages of the K1 family, infecting E. coli K1, specifically recognize and degrade this polySia capsule utilizing tailspike endosialidases. While the crystal structure for the catalytic domain of the endosialidase of bacteriophage K1F (endoNF) has been solved, there is yet no structural information on the mode of polySia binding and cleavage available. The crystal structure of activity deficient active-site mutants of the homotrimeric endoNF cocrystallized with oligomeric sialic acid identified three independent polySia binding sites in each endoNF monomer. The bound oligomeric sialic acid displays distinct conformations at each site. In the active site, a Sia(3) molecule is bound in an extended conformation representing the enzyme-product complex. Structural and biochemical data supported by molecular modeling enable to propose a reaction mechanism for polySia cleavage by endoNF.

• Authors: Eike Christian Schulz, David Schwarzer, Martin Frank, Katharina Stummeyer, Martina Mühlenhoff, Achim Dickmanns, Rita Gerardy-Schahn, Ralf Ficner
• Journal: Journal of molecular biology (J Mol Biol) Vol. 397 Issue 1 Pg. 341-51 (Mar 19 2010) ISSN: 1089-8638 [Electronic] Netherlands
• PMID: 20096705 (Publication Type: Journal Article)
• Copyright: Copyright (c) 2010 Elsevier Ltd. All rights reserved.

Chemical References

• Mutant Proteins
• Sialic Acids
• Viral Tail Proteins
• polysialic acid
• Glycoside Hydrolases
• endo-N-acetylneuraminidase
• tailspike protein, bacteriophage
• Neuraminidase

Topics

• Bacteriophages (enzymology)
• Catalytic Domain
• Crystallography, X-Ray
• Glycoside Hydrolases
• Models, Molecular
• Mutant Proteins (chemistry, metabolism)
• Neuraminidase (chemistry, metabolism)
• Protein Structure, Secondary
• Sialic Acids (metabolism)
• Substrate Specificity
• Viral Tail Proteins (chemistry, metabolism)