TY - JOUR

T1 - Electrochemical Single‐Molecule AFM of the Redox Metalloenzyme Copper Nitrite Reductase in Action

A1 - Hao,Xian

A1 - Zhang,Jingdong

A1 - Christensen,Hans Erik Mølager

A1 - Wang,Hongda

A1 - Ulstrup,Jens

AU - Hao,Xian

AU - Zhang,Jingdong

AU - Christensen,Hans Erik Mølager

AU - Wang,Hongda

AU - Ulstrup,Jens

PB - Wiley - V C H Verlag GmbH & Co. KGaA

PY - 2012

Y1 - 2012

N2 - We studied the electrochemical behavior of the redox metalloenzyme copper nitrite reductase (CNiR, Achromobacter xylosoxidans) immobilized on a Au(111)‐electrode surface modified by a self‐assembled cysteamine molecular monolayer (SAM) using a combination of cyclic voltammetry and electrochemically‐controlled atomic force microscopy (in situ AFM). The enzyme showed no voltammetric signals in the absence of nitrite substrate, whereas a strong reductive electrocatalytic signal appeared in the presence of nitrite. Such a pattern is common in protein film and monolayer voltammetry and points to conformational changes in the enzyme upon substrate binding. Binding thus either improves the enzyme/electrode contact, or opens intramolecular electron‐transfer channels between the redox center for electron inlet (a type I copper center) and the catalytic site for nitrite reduction (a type II copper center). The in situ AFM data are at the level of the single CuNiR enzyme molecule. The voltammetric patterns were paralleled by a clear increase (swelling) of the molecular height when the electrochemical potential traversed the region from resting to the electrocatalytically active redox enzyme function in the presence of nitrite. No change in size was observed in the absence of nitrite over the same potential range. The enzyme size variation is suggested to offer clues to the broadly observed substrate triggering in metalloenzyme monolayer voltammetry.

AB - We studied the electrochemical behavior of the redox metalloenzyme copper nitrite reductase (CNiR, Achromobacter xylosoxidans) immobilized on a Au(111)‐electrode surface modified by a self‐assembled cysteamine molecular monolayer (SAM) using a combination of cyclic voltammetry and electrochemically‐controlled atomic force microscopy (in situ AFM). The enzyme showed no voltammetric signals in the absence of nitrite substrate, whereas a strong reductive electrocatalytic signal appeared in the presence of nitrite. Such a pattern is common in protein film and monolayer voltammetry and points to conformational changes in the enzyme upon substrate binding. Binding thus either improves the enzyme/electrode contact, or opens intramolecular electron‐transfer channels between the redox center for electron inlet (a type I copper center) and the catalytic site for nitrite reduction (a type II copper center). The in situ AFM data are at the level of the single CuNiR enzyme molecule. The voltammetric patterns were paralleled by a clear increase (swelling) of the molecular height when the electrochemical potential traversed the region from resting to the electrocatalytically active redox enzyme function in the presence of nitrite. No change in size was observed in the absence of nitrite over the same potential range. The enzyme size variation is suggested to offer clues to the broadly observed substrate triggering in metalloenzyme monolayer voltammetry.

U2 - 10.1002/cphc.201200220

DO - 10.1002/cphc.201200220

JO - ChemPhysChem

JF - ChemPhysChem

SN - 1439-4235

IS - 12

VL - 13

SP - 2919

EP - 2924

ER -