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- Broeker, N. K. (author)
Single amino acid exchange in bacteriophage HK620 tailspike protein results in thousand-fold increase of its oligosaccharide affinity
- Article/chapterEnglish2013
Publisher, publication year, extent ...
- 2012-08-24
- Oxford University Press (OUP),2013
- printrdacarrier
Numbers
- LIBRIS-ID:oai:DiVA.org:uu-192025
- https://urn.kb.se/resolve?urn=urn:nbn:se:uu:diva-192025URI
- https://doi.org/10.1093/glycob/cws126DOI
Supplementary language notes
- Language:English
- Summary in:English
Part of subdatabase
- SwePubSwePub
Classification
Notes
- 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
- bacterial O-antigen
- carbohydrate interaction
- site-directed mutagenesis
- structural thermodynamics
- tailspike protein
- bacteriophage hk620 tailspike protein
- carbohydrate
- glutamic acid
- glutamine
- mutant protein
- O antigen
- oligosaccharide
- solvent
- unclassified drug
- virus protein
- amino acid substitution
- article
- bacteriophage
- bacteriophage hk620
- binding site
- complex formation
- crystal structure
- dissociation constant
- enthalpy
- Escherichia coli
- heat
- hydrogen bond
- hydrophobicity
- isothermal titration calorimetry
- nonhuman
- priority journal
- protein binding
- reaction analysis
- room temperature
Added entries (persons, corporate bodies, meetings, titles ...)
- Gohlke, U. (author)
- Müller, J. J. (author)
- Uetrecht, CharlotteUppsala universitet,Molekylär biofysik(Swepub:uu)chaue348 (author)
- Heinemann, U. (author)
- Seckler, R. (author)
- Barbirz, S. (author)
- Uppsala universitetMolekylär biofysik (creator_code:org_t)
Related titles
- In:Glycobiology: Oxford University Press (OUP)23:1, s. 59-680959-66581460-2423
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