Enzyme electrophoresis is the separation of enzymes by their movement in an electric field through a supporting medium. Gel-based separation produces spatially resolved protein bands that can be examined for catalytic activity. In gel zymography, substrate conversion generates a localized signal corresponding to an active enzyme form [1]. Native electrophoresis separates proteins under conditions that can preserve folded structures or associated molecular components [2]. Combining native separation with an in-gel activity reaction links a resolved complex to its catalytic function [3]. Electrophoretic patterns may also represent variation among enzyme forms across multiple loci [4]. The observed pattern reflects protein migration, the separation matrix, the biochemical conditions maintained during electrophoresis, and the specificity of the subsequent activity-detection reaction [5].
- electrophoresis
- zymography
- enzyme activity
- protein separation
Enzyme Production and Characterization·Biotechnology·Biochemistry, Genetics and Molecular Biology·Life Sciences
References
- Jennifer Vandooren; Nathalie Geurts; Erik Martens; Philippe E Van Den Steen; Ghislain Opdenakker; Zymography methods for visualizing hydrolytic enzymes. Nat. Methods 2013, 10, 211-220, 10.1038/nmeth.2371.
- Hermann Schägger; Gebhard von Jagow; Blue native electrophoresis for isolation of membrane protein complexes in enzymatically active form. Anal. Biochem. 1991, 199, 223-231, 10.1016/0003-2697(91)90094-a.
- Laura Gardano; Linda Holland; Rena Oulton; Thierry Le Bihan; Lea Harrington; Native gel electrophoresis of human telomerase distinguishes active complexes with or without dyskerin. Nucleic Acids Res. 2011, 40, e36-e36, 10.1093/nar/gkr1243.
- Gan Wang; Thomas S. Whittam; Claire M. Berg; Douglas E. Berg; RAPD (arbitrary primer) PCR is more sensitive than multilocus enzyme electrophoresis for distinguishing related bacterial strains. Nucleic Acids Res. 1993, 21, 5930-5933, 10.1093/nar/21.25.5930.
- Thierry Rabilloud; Variations on a theme: Changes to electrophoretic separations that can make a difference. J. Proteom. 2010, 73, 1562-1572, 10.1016/j.jprot.2010.04.001.