Engineering a disulfide-gated switch in streptavidin enables reversible binding without sacrificing binding affinity
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 koff, the disulfide in M88 stabilizes the closed conformation, decreasing koff 260-fold relative to streptavidin. The simple and efficient reduction of this disulfide increases koff 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.
- Authors:
-
- Univ. of Calgary, AB (Canada). Dept. of Biological Scienes
- Univ. of Calgary, AB (Canada). Dept. of Biological Scienes; Univ. of Hawaii, Honolulu, HI (United States). Dept. of Molecular Biosciences and Bioengineering
- Publication Date:
- Research Org.:
- SLAC National Accelerator Lab., Menlo Park, CA (United States)
- Sponsoring Org.:
- USDOE Office of Science (SC), Basic Energy Sciences (BES). Scientific User Facilities Division; Natural Sciences and Engineering Research Council of Canada (NSERC); National Institutes of Health (NIH); National Center for Research Resources
- OSTI Identifier:
- 1816245
- Grant/Contract Number:
- AC02-76SF00515; 690573; 05728; P41GM103393; P41RR001209
- Resource Type:
- Accepted Manuscript
- Journal Name:
- Scientific Reports
- Additional Journal Information:
- Journal Volume: 10; Journal Issue: 1; Journal ID: ISSN 2045-2322
- Publisher:
- Nature Publishing Group
- Country of Publication:
- United States
- Language:
- English
- Subject:
- 59 BASIC BIOLOGICAL SCIENCES; biochemistry; biological techniques; molecular engineering; protein design; proteins
Citation Formats
Marangoni, Jesse M., Wu, Sau-Ching, Fogen, Dawson, Wong, Sui-Lam, and Ng, Kenneth K. S. Engineering a disulfide-gated switch in streptavidin enables reversible binding without sacrificing binding affinity. United States: N. p., 2020.
Web. doi:10.1038/s41598-020-69357-5.
Marangoni, Jesse M., Wu, Sau-Ching, Fogen, Dawson, Wong, Sui-Lam, & Ng, Kenneth K. S. Engineering a disulfide-gated switch in streptavidin enables reversible binding without sacrificing binding affinity. United States. https://doi.org/10.1038/s41598-020-69357-5
Marangoni, Jesse M., Wu, Sau-Ching, Fogen, Dawson, Wong, Sui-Lam, and Ng, Kenneth K. S. Mon .
"Engineering a disulfide-gated switch in streptavidin enables reversible binding without sacrificing binding affinity". United States. https://doi.org/10.1038/s41598-020-69357-5. https://www.osti.gov/servlets/purl/1816245.
@article{osti_1816245,
title = {Engineering a disulfide-gated switch in streptavidin enables reversible binding without sacrificing binding affinity},
author = {Marangoni, Jesse M. and Wu, Sau-Ching and Fogen, Dawson and Wong, Sui-Lam and Ng, Kenneth K. S.},
abstractNote = {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 koff, the disulfide in M88 stabilizes the closed conformation, decreasing koff 260-fold relative to streptavidin. The simple and efficient reduction of this disulfide increases koff 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.},
doi = {10.1038/s41598-020-69357-5},
journal = {Scientific Reports},
number = 1,
volume = 10,
place = {United States},
year = {Mon Jul 27 00:00:00 EDT 2020},
month = {Mon Jul 27 00:00:00 EDT 2020}
}
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