TY  - JOUR
AU  - Ataka, K.
AU  - Heberle, J.
TI  - Use of Surface Enhanced Infrared Absorption Spectroscopy (SEIRA) to probe the functionality of a protein monolayer
JO  - Biopolymers
VL  - 82
SN  - 0006-3525
CY  - New York, NY
PB  - Wiley
M1  - PreJuSER-57463
SP  - 415 - 419
PY  - 2006
N1  - Record converted from VDB: 12.11.2012
AB  - We present a novel infrared method to investigate the functionality of a protein monolayer tethered to a metal substrate. The approach employs Surface Enhanced Infrared Absorption Spectroscopy (SEIRAS), which renders high surface sensitivity by enhancing the signal of the adsorbed protein by up to approximately 2 orders of magnitude. We demonstrate that the electrochemically induced absorption changes of a cytochrome c monolayer can be observed with excellent signal-to-noise ratio when the protein is adhered to a modified gold surface. To probe membrane proteins, a concept is introduced for the oriented incorporation into solid supported lipid bilayers. Recombinant cytochrome c oxidase solubilized in detergent is immobilized on a chemically modified gold surface via the affinity of its histidine (His)-tag to a nickel-chelating nitro-triacetic acid (NTA) surface. The protein monolayer is reconstituted into the lipid environment by detergent removal. Changing the orientation of the protein with respect to the metal surface is achieved by inserting the His-tag on either side of the membrane protein surface. Orientational control is mandatory for experiments in which electrons are injected from the electrode into the protein. The presented methodology opens new avenues to study the mechanism of the biomedically relevant class of electron and voltage-gated proteins on the atomic level.
KW  - Electrochemistry: instrumentation
KW  - Electrochemistry: methods
KW  - Electrodes
KW  - Electron Transport Complex IV: chemistry
KW  - Electron Transport Complex IV: metabolism
KW  - Gold: chemistry
KW  - Membrane Proteins: chemistry
KW  - Membrane Proteins: metabolism
KW  - Models, Molecular
KW  - Proteins: chemistry
KW  - Proteins: metabolism
KW  - Spectrophotometry, Infrared: instrumentation
KW  - Spectrophotometry, Infrared: methods
KW  - Surface Properties
KW  - Time Factors
KW  - Membrane Proteins (NLM Chemicals)
KW  - Proteins (NLM Chemicals)
KW  - Gold (NLM Chemicals)
KW  - Electron Transport Complex IV (NLM Chemicals)
KW  - J (WoSType)
LB  - PUB:(DE-HGF)16
C6  - pmid:16518850
UR  - <Go to ISI:>//WOS:000238241900027
DO  - DOI:10.1002/bip.20501
UR  - https://juser.fz-juelich.de/record/57463
ER  -