Engineering a disulfide-gated switch in streptavidin enables reversible binding without sacrificing binding affinity

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...

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Bibliographic Details
Published in:Scientific reports 2020-07, Vol.10 (1), p.12483-12483, Article 12483
Main Authors: Marangoni, Jesse M., Wu, Sau-Ching, Fogen, Dawson, Wong, Sui-Lam, Ng, Kenneth K. S.
Format: Article
Language:English
Subjects:
631/1647
631/1647/338
631/1647/338/469
631/45
631/45/612
Acids
Affinity
BASIC BIOLOGICAL SCIENCES
Binding Sites
biochemistry
biological techniques
Biotin
Biotin - chemistry
Chemical bonds
Conformation
Crystallography, X-Ray
Disulfide bonds
Disulfides - chemistry
Engineering
Fluorescein - chemistry
Humanities and Social Sciences
Kinetics
Ligands
Models, Molecular
molecular engineering
multidisciplinary
Mutant Proteins - chemistry
Mutation
Oxidation-Reduction
Peptides
Protein Binding
protein design
Protein Engineering
Protein Stability
Proteins
Science
Science (multidisciplinary)
Streptavidin
Streptavidin - chemistry
Structure-function relationships
Temperature
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Summary: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 off , the disulfide in M88 stabilizes the closed conformation, decreasing k off 260-fold relative to streptavidin. The simple and efficient reduction of this disulfide increases k off 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.
ISSN:2045-2322
2045-2322
DOI:10.1038/s41598-020-69357-5