4AVZ

Tailspike protein mutant E372Q of E. coli bacteriophage HK620

4AVZ の概要

• エントリーDOI: 10.2210/pdb4avz/pdb
• 関連するPDBエントリー: 2VJI 2VJJ 2X6W 2X6X 2X6Y 2X85
• 分子名称: TAIL SPIKE PROTEIN, TRIS-HYDROXYMETHYL-METHYL-AMMONIUM (3 entities in total)
• 機能のキーワード: viral protein, endo-n-acetylglucosaminidase, o-antigen
• 由来する生物種: SALMONELLA PHAGE HK620
• タンパク質・核酸の鎖数: 1
• 化学式量合計: 64826.51

構造登録者

Gohlke, U.,Broeker, N.K.,Mueller, J.J.,Uetrecht, C.,Heinemann, U.,Seckler, R.,Barbirz, S. (登録日: 2012-05-30, 公開日: 2012-09-26, 最終更新日: 2023-12-20)

主引用文献

Broeker, N.K.,Gohlke, U.,Muller, J.J.,Uetrecht, C.,Heinemann, U.,Seckler, R.,Barbirz, S.
Single Amino Acid Exchange in Bacteriophage Hk620 Tailspike Protein Results in Thousand-Fold Increase of its Oligosaccharide Affinity.
Glycobiology, 23:59-, 2013

PubMed Abstract: 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.

PubMed: 22923442
DOI: 10.1093/GLYCOB/CWS126

実験手法

X-RAY DIFFRACTION (1.82 Å)