Controlled levels of protein modification through a chromatography-mediated bioconjugation

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 describ...

Full description

Saved in:
Bibliographic Details
Published in:Chemical science (Cambridge) 2015-04, Vol.6 (4), p.2596-2601
Main Authors: Kwant, Richard L, Jaffe, Jake, Palmere, Peter J, Francis, Matthew B
Format: Article
Language:English
Subjects:
BASIC BIOLOGICAL SCIENCES
Chemical reactions
Chromatography
Construction
Functional groups
Handles
INORGANIC, ORGANIC, PHYSICAL, AND ANALYTICAL CHEMISTRY
Proteins
Scaffolds
Citations: Items that this one cites
Items that cite this one
Online Access:Get full text
Tags: Add Tag
No Tags, Be the first to tag this record!
Description
Summary: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 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.
ISSN:2041-6520
2041-6539
DOI:10.1039/c4sc03790a