Selenomethionine as an expressible handle for bioconjugations
Abstract
Site-selective chemical bioconjugation reactions are enabling tools for the chemical biologist. Guided by a careful study of the selenomethionine (SeM) benzylation, we have refined the reaction to meet the requirements of practical protein bioconjugation. SeM is readily introduced through auxotrophic expression and exhibits unique nucleophilic properties that allow it to be selectively modified even in the presence of cysteine. The resulting benzylselenonium adduct is stable at physiological pH, is selectively labile to glutathione, and embodies a broadly tunable cleavage profile. Specifically, a 4-bromomethylphenylacetyl (BrMePAA) linker has been applied for efficient conjugation of complex organic molecules to SeM-containing proteins. This expansion of the bioconjugation toolkit has broad potential in the development of chemically enhanced proteins.
Additional Information
© 2021 National Academy of Sciences. Published under the PNAS license. Edited by David Baker, University of Washington, Seattle, WA, and approved January 4, 2021 (received for review March 24, 2020). Financial support for this work was from NIH (GM-132787 to P.E.D. and GM092740 to T.O.). D.T.F. was supported by the National Center for Advancing Translational Sciences, NIH, through Grant UL1 TR002551 and linked award TL1 TR002551. The content is solely the responsibility of the authors and does not necessarily represent the official views of the NIH. We are grateful to Dr. Dee-Hua Huang and Dr. Laura Pasternack (Scripps Research) for assistance with nuclear magnetic resonance spectroscopy. Data Availability: All study data are included in this article and/or SI Appendix. Author contributions: D.T.F., D.E.H., and P.E.D. designed research; D.T.F., J.C.J.H., K.W.K., D.E.H., C.L., P.A.C., and T.O. performed research; D.T.F. and J.S.C. contributed new reagents/analytic tools; D.T.F., J.C.J.H., D.E.H., T.O., and P.E.D. analyzed data; and D.T.F. and P.E.D. wrote the paper. The authors declare no competing interest. This article is a PNAS Direct Submission. This article contains supporting information online at https://www.pnas.org/lookup/suppl/doi:10.1073/pnas.2005164118/-/DCSupplemental.Attached Files
Published - e2005164118.full.pdf
Supplemental Material - pnas.2005164118.sapp.pdf
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Additional details
- PMCID
- PMC7923357
- Eprint ID
- 108120
- Resolver ID
- CaltechAUTHORS:20210219-111631143
- GM-132787
- NIH
- GM092740
- NIH
- UL1 TR002551
- NIH
- TL1 TR002551
- NIH
- Created
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2021-02-19Created from EPrint's datestamp field
- Updated
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2023-07-17Created from EPrint's last_modified field