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Updated: Jul 11, 2026

Proteomics to Identify Proteins Interacting with P2X2 Ligand-Gated Cation Channels
Published on: May 19, 2009
Real-time kinetic analysis of limited proteolysis by ion-exchange chromatography using compressed, non-porous agarose
J Mohammad1, K O Eriksson, G Johansson
1Department of Biochemistry, University of Uppsala, Sweden.
Abstract:
In biotechnology there is a great need for methods that can be used for kinetic studies of ongoing processes, such as limited proteolysis. Ion-exchange chromatography on a nonporous, compressed agarose matrix was used in the study presented in this paper to monitor papain cleavage of fungal cellulases. This non-porous matrix allows completion of a chromatographic analysis, with baseline resolution, within 30-60 s. Utilizing this very fast chromatographic method one can monitor a proteolysis 'continuously' by repeated chromatographic analysis of the incubation mixture and, if desired, terminate it at the optimal moment. Fractions were also collected for identification by activity assays and analytical SDS-PAGE.
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