Issue 4, 2015

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.

Graphical abstract: Controlled levels of protein modification through a chromatography-mediated bioconjugation

Supplementary files

Article information

Article type
Edge Article
Submitted
08 Dec 2014
Accepted
30 Jan 2015
First published
27 Feb 2015
This article is Open Access

All publication charges for this article have been paid for by the Royal Society of Chemistry
Creative Commons BY license

Chem. Sci., 2015,6, 2596-2601

Author version available

Controlled levels of protein modification through a chromatography-mediated bioconjugation

R. L. Kwant, J. Jaffe, P. J. Palmere and M. B. Francis, Chem. Sci., 2015, 6, 2596 DOI: 10.1039/C4SC03790A

This article is licensed under a Creative Commons Attribution 3.0 Unported Licence. You can use material from this article in other publications without requesting further permissions from the RSC, provided that the correct acknowledgement is given.

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