6VJL

Streptavidin mutant M112 (G26C/A46C)

6VJL の概要

• エントリーDOI: 10.2210/pdb6vjl/pdb
• 分子名称: Streptavidin, BIOTIN (3 entities in total)
• 機能のキーワード: biotin-binding protein
• 由来する生物種: Streptomyces avidinii
• タンパク質・核酸の鎖数: 1
• 化学式量合計: 16958.52

構造登録者

Marangoni, J.M.,Wu, S.C.,Fogen, D.,Wong, S.L.,Ng, K.K.S. (登録日: 2020-01-16, 公開日: 2020-12-23, 最終更新日: 2024-11-06)

主引用文献

Marangoni, J.M.,Wu, S.C.,Fogen, D.,Wong, S.L.,Ng, K.K.S.
Engineering a disulfide-gated switch in streptavidin enables reversible binding without sacrificing binding affinity.
Sci Rep, 10:12483-12483, 2020

PubMed Abstract: Although high affinity binding between streptavidin and biotin is widely exploited, the accompanying low rate of dissociation prevents its use in many applications where rapid ligand release is also required. To combine extremely tight and reversible binding, we have introduced disulfide bonds into opposite sides of a flexible loop critical for biotin binding, creating streptavidin muteins (M88 and M112) with novel disulfide-switchable binding properties. Crystal structures reveal how each disulfide exerts opposing effects on structure and function. Whereas the disulfide in M112 disrupts the closed conformation to increase k, the disulfide in M88 stabilizes the closed conformation, decreasing k 260-fold relative to streptavidin. The simple and efficient reduction of this disulfide increases k 19,000-fold, thus creating a reversible redox-dependent switch with 70-fold faster dissociation kinetics than streptavidin. The facile control of disulfide formation in M88 will enable the development of many new applications requiring high affinity and reversible binding.

PubMed: 32719366
DOI: 10.1038/s41598-020-69357-5

実験手法

X-RAY DIFFRACTION (1.3 Å)