Related Experiment Videos
Site-specific introduction of an electroactive label into a non-electroactive enzyme (beta-lactamase I)
K Di Gleria1, C M Halliwell, C Jacob
1New Chemistry Laboratory, University of Oxford, UK.
FEBS Letters
|January 3, 1997
Abstract:
A cysteine residue was introduced close to the active site of beta-lactamase I by site-directed mutagenesis to replace tyrosine-105 and was subsequently modified with an electroactive SH-specific reagent, N-(2-ferrocene-ethyl)maleimide. The resulting modified enzyme became electroactive, showing good quasireversible electrochemistry which was characteristic of the attached ferrocene moiety while retaining its specific enzymatic activity. In the presence of a suicide substrate, 6beta-iodopenicillanic acid, the redox potential shifted +20 mV suggesting that the label was sensitive to changes in the active site of the enzyme.