Controlled levels of protein modification through a chromatography-mediated bioconjugation
Abstract
Synthetically modified proteins are increasingly finding applications as well-defined scaffolds for materials. In practice it remains difficult to construct bioconjugates with precise levels of modification because of the limited number of repeated functional groups on proteins. This article describes a method to control the level of protein modification in cases where there exist multiple potential modification sites. A protein is first tagged with a handle using any of a variety of modification chemistries. This handle is used to isolate proteins with a particular number of modifications via affinity chromatography, and then the handle is elaborated with a desired moiety using an oxidative coupling reaction. This method results in a sample of protein with a well-defined number of modifications, and we find it particularly applicable to systems like protein homomultimers in which there is no way to discern between chemically identical subunits. We demonstrate the use of this method in the construction of a protein-templated light-harvesting mimic, a type of system which has historically been difficult to make in a well-defined manner.
- Authors:
-
- Univ. of California, Berkeley, CA (United States). Dept. of Chemistry.
- Univ. of California, Berkeley, CA (United States). Dept. of Chemistry; Lawrence Berkeley National Lab., Berkeley, CA (United States). Materials Sciences Div.
- Publication Date:
- Research Org.:
- Lawrence Berkeley National Lab. (LBNL), Berkeley, CA (United States)
- Sponsoring Org.:
- USDOE Advanced Research Projects Agency - Energy (ARPA-E)
- OSTI Identifier:
- 1214442
- Grant/Contract Number:
- AC02-05CH11231
- Resource Type:
- Accepted Manuscript
- Journal Name:
- Chemical Science
- Additional Journal Information:
- Journal Volume: 6; Journal Issue: 4; Journal ID: ISSN 2041-6520
- Publisher:
- Royal Society of Chemistry
- Country of Publication:
- United States
- Language:
- English
- Subject:
- 37 INORGANIC, ORGANIC, PHYSICAL, AND ANALYTICAL CHEMISTRY; 59 BASIC BIOLOGICAL SCIENCES
Citation Formats
Kwant, Richard L., Jaffe, Jake, Palmere, Peter J., and Francis, Matthew B. Controlled levels of protein modification through a chromatography-mediated bioconjugation. United States: N. p., 2015.
Web. doi:10.1039/C4SC03790A.
Kwant, Richard L., Jaffe, Jake, Palmere, Peter J., & Francis, Matthew B. Controlled levels of protein modification through a chromatography-mediated bioconjugation. United States. https://doi.org/10.1039/C4SC03790A
Kwant, Richard L., Jaffe, Jake, Palmere, Peter J., and Francis, Matthew B. Fri .
"Controlled levels of protein modification through a chromatography-mediated bioconjugation". United States. https://doi.org/10.1039/C4SC03790A. https://www.osti.gov/servlets/purl/1214442.
@article{osti_1214442,
title = {Controlled levels of protein modification through a chromatography-mediated bioconjugation},
author = {Kwant, Richard L. and Jaffe, Jake and Palmere, Peter J. and Francis, Matthew B.},
abstractNote = {Synthetically modified proteins are increasingly finding applications as well-defined scaffolds for materials. In practice it remains difficult to construct bioconjugates with precise levels of modification because of the limited number of repeated functional groups on proteins. This article describes a method to control the level of protein modification in cases where there exist multiple potential modification sites. A protein is first tagged with a handle using any of a variety of modification chemistries. This handle is used to isolate proteins with a particular number of modifications via affinity chromatography, and then the handle is elaborated with a desired moiety using an oxidative coupling reaction. This method results in a sample of protein with a well-defined number of modifications, and we find it particularly applicable to systems like protein homomultimers in which there is no way to discern between chemically identical subunits. We demonstrate the use of this method in the construction of a protein-templated light-harvesting mimic, a type of system which has historically been difficult to make in a well-defined manner.},
doi = {10.1039/C4SC03790A},
journal = {Chemical Science},
number = 4,
volume = 6,
place = {United States},
year = {Fri Feb 27 00:00:00 EST 2015},
month = {Fri Feb 27 00:00:00 EST 2015}
}
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