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Single amino acid e...
Single amino acid exchange in bacteriophage HK620 tailspike protein results in thousand-fold increase of its oligosaccharide affinity
- Article/chapterEnglish2013
Publisher, publication year, extent ...
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2012-08-24
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Oxford University Press (OUP),2013
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printrdacarrier
Numbers
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LIBRIS-ID:oai:DiVA.org:uu-192025
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https://urn.kb.se/resolve?urn=urn:nbn:se:uu:diva-192025URI
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https://doi.org/10.1093/glycob/cws126DOI
Supplementary language notes
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Language:English
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Summary in:English
Part of subdatabase
Classification
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Subject category:ref swepub-contenttype
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Subject category:art swepub-publicationtype
Notes
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Bacteriophage HK620 recognizes and cleaves the O-antigen polysaccharide of Escherichia coli serogroup O18A1 with its tailspike protein (TSP). HK620TSP binds hexasaccharide fragments with low affinity, but single amino acid exchanges generated a set of high-affinity mutants with submicromolar dissociation constants. Isothermal titration calorimetry showed that only small amounts of heat were released upon complex formation via a large number of direct and solvent-mediated hydrogen bonds between carbohydrate and protein. At room temperature, association was both enthalpy- and entropy-driven emphasizing major solvent rearrangements upon complex formation. Crystal structure analysis showed identical protein and sugar conformers in the TSP complexes regardless of their hexasaccharide affinity. Only in one case, a TSP mutant bound a different hexasaccharide conformer. The extended sugar binding site could be dissected in two regions: first, a hydrophobic pocket at the reducing end with minor affinity contributions. Access to this site could be blocked by a single aspartate to asparagine exchange without major loss in hexasaccharide affinity. Second, a region where the specific exchange of glutamate for glutamine created a site for an additional water molecule. Side-chain rearrangements upon sugar binding led to desolvation and additional hydrogen bonding which define this region of the binding site as the high-affinity scaffold.
Subject headings and genre
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bacterial O-antigen
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carbohydrate interaction
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site-directed mutagenesis
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structural thermodynamics
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tailspike protein
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bacteriophage hk620 tailspike protein
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carbohydrate
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glutamic acid
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glutamine
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mutant protein
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O antigen
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oligosaccharide
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solvent
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unclassified drug
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virus protein
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amino acid substitution
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article
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bacteriophage
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bacteriophage hk620
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binding site
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complex formation
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crystal structure
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dissociation constant
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enthalpy
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Escherichia coli
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heat
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hydrogen bond
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hydrophobicity
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isothermal titration calorimetry
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nonhuman
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priority journal
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protein binding
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reaction analysis
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room temperature
Added entries (persons, corporate bodies, meetings, titles ...)
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Gohlke, U.
(author)
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Müller, J. J.
(author)
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Uetrecht, CharlotteUppsala universitet,Molekylär biofysik(Swepub:uu)chaue348
(author)
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Heinemann, U.
(author)
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Seckler, R.
(author)
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Barbirz, S.
(author)
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Uppsala universitetMolekylär biofysik
(creator_code:org_t)
Related titles
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In:Glycobiology: Oxford University Press (OUP)23:1, s. 59-680959-66581460-2423
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