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Scanning Electrochemical Microscopy of Cytochrome c Peroxidase through the Orientation‐Controlled Immobilisation of Cytochrome c
Author(s) -
Gunawan Christian A.,
Nam Ekaterina V.,
Marquis Christopher P.,
Gooding J. Justin,
Thordarson Pall,
Zhao Chuan
Publication year - 2016
Publication title -
chemelectrochem
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.182
H-Index - 59
ISSN - 2196-0216
DOI - 10.1002/celc.201600195
Subject(s) - chemistry , cytochrome c , maleimide , peroxidase , cytochrome c peroxidase , docking (animal) , redox , enzyme , electrochemistry , combinatorial chemistry , stereochemistry , biochemistry , organic chemistry , mitochondrion , electrode , medicine , nursing
Immobilisation of enzymes onto surfaces with defined orientation is of particular importance in order to optimise their activity and capacity to dock electron‐transfer partners. Local characterisation methods are thus crucial to understanding the activity–orientation relationship of immobilised enzymes for long‐term storage and applications. Herein, cytochrome c (cyt c ) is chemoselectively immobilised through a cysteine functional group with a maleimide‐functionalised gold surface, thus controlling cyt c orientation. X‐ray photoelectron spectroscopy confirmed successful cyt c immobilisation. Scanning electrochemical microscopy methodologies were developed to quantify the activity of immobilised cyt c and cytochrome c peroxidase (C c P). The results suggest optimal enzymatic activities and docking ability for surface‐tethered cyt c are obtained through the cysteine–maleimide linkage, in comparison to conventional indiscriminate lysine–carboxyl linkages. This study reveals the strong influence of cyt c orientation, as a result of linking strategy, upon the docking ability of redox‐electron partner C c P.