Use of surface enhanced infrared absorption spectroscopy (SEIRA) to probe the functionality of a protein monolayer

Biopolymers
Kenichi Ataka, Joachim Heberle

Abstract

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 p...Continue Reading

References

Apr 2, 1999·Biosensors & Bioelectronics·D KrögerH Vogel
Apr 24, 2003·Journal of the American Chemical Society·Kenichi Ataka, Joachim Heberle
Sep 2, 2004·Biophysical Journal·Frank GiessWolfgang Knoll
Sep 7, 2005·Langmuir : the ACS Journal of Surfaces and Colloids·Dirk MayerAndreas Offenhäusser

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Citations

Aug 23, 2008·Proceedings of the National Academy of Sciences of the United States of America·X JiangJ Heberle
Feb 18, 2016·Analytical and Bioanalytical Chemistry·Ángela I López-Lorente, Boris Mizaikoff
Jun 19, 2012·Chemphyschem : a European Journal of Chemical Physics and Physical Chemistry·Philipp PinkerneilCarsten Kötting
Mar 6, 2009·Mass Spectrometry Reviews·Jamie D DunnMerlin L Bruening
Aug 10, 2013·Biochimica Et Biophysica Acta·Stefanie E GlassfordSergei G Kazarian
Nov 8, 2011·Chemical Communications : Chem Comm·Philip A Ash, Kylie A Vincent
Jul 9, 2016·Langmuir : the ACS Journal of Surfaces and Colloids·Mohammad A FallahKarin Hauser
Dec 20, 2008·Applied Spectroscopy·Sara A LoveChristy L Haynes
Feb 6, 2017·Proceedings of the National Academy of Sciences of the United States of America·Natalia GrytsykPetra Hellwig
Feb 23, 2019·Proceedings of the National Academy of Sciences of the United States of America·Natalia GrytsykPetra Hellwig
Aug 22, 2020·Chemical Reviews·Frederic Melin, Petra Hellwig
Apr 25, 2019·Biochemistry·Alejandra de Miguel CatalinaMaria Andrea Mroginski

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