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Cyanylated Cysteine: A Covalently Attached Vibrational Probe of Protein−Lipid Contacts

[Image: see text] Cyanylated cysteine, or β-thiocyanatoalanine, is an artificial amino acid that can be introduced into peptides and proteins by post-translational chemical modification of solvent-exposed cysteine side chains, and thus it can be used in any protein with a suitable expression and mut...

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Autores principales: McMahon, Heather A., Alfieri, Katherine N., Clark, Katherine A. A., Londergan, Casey H.
Formato: Texto
Lenguaje:English
Publicado: American Chemical Society 2010
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2836368/
https://www.ncbi.nlm.nih.gov/pubmed/20228945
http://dx.doi.org/10.1021/jz1000177
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author McMahon, Heather A.
Alfieri, Katherine N.
Clark, Katherine A. A.
Londergan, Casey H.
author_facet McMahon, Heather A.
Alfieri, Katherine N.
Clark, Katherine A. A.
Londergan, Casey H.
author_sort McMahon, Heather A.
collection PubMed
description [Image: see text] Cyanylated cysteine, or β-thiocyanatoalanine, is an artificial amino acid that can be introduced into peptides and proteins by post-translational chemical modification of solvent-exposed cysteine side chains, and thus it can be used in any protein with a suitable expression and mutagenesis system. In this study, cyanylated cysteine is introduced at selected sites in two model peptides that have been shown to bind to membrane interfaces: a membrane-binding sequence of the human myelin basic protein and the antimicrobial peptide CM15. Far-UV circular dichroism indicates that the secondary structures of the bound peptides are not influenced by introduction of the artificial side chain. Infrared spectra of both systems in buffer and exposed to dodecylphosphocholine micelles indicate that the CN stretching absorption band of cyanylated cysteine can clearly distinguish between membrane burial and solvent exposure of the artificial side chain. Since infrared spectroscopy can be applied in a wide variety of lipid systems, and since cyanylated cysteine can be introduced into proteins of arbitrary size via mutagenesis and post-translational modification, this new probe could see wide use in characterizing the protein−lipid interactions of membrane proteins.
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spelling pubmed-28363682010-03-11 Cyanylated Cysteine: A Covalently Attached Vibrational Probe of Protein−Lipid Contacts McMahon, Heather A. Alfieri, Katherine N. Clark, Katherine A. A. Londergan, Casey H. J Phys Chem Lett [Image: see text] Cyanylated cysteine, or β-thiocyanatoalanine, is an artificial amino acid that can be introduced into peptides and proteins by post-translational chemical modification of solvent-exposed cysteine side chains, and thus it can be used in any protein with a suitable expression and mutagenesis system. In this study, cyanylated cysteine is introduced at selected sites in two model peptides that have been shown to bind to membrane interfaces: a membrane-binding sequence of the human myelin basic protein and the antimicrobial peptide CM15. Far-UV circular dichroism indicates that the secondary structures of the bound peptides are not influenced by introduction of the artificial side chain. Infrared spectra of both systems in buffer and exposed to dodecylphosphocholine micelles indicate that the CN stretching absorption band of cyanylated cysteine can clearly distinguish between membrane burial and solvent exposure of the artificial side chain. Since infrared spectroscopy can be applied in a wide variety of lipid systems, and since cyanylated cysteine can be introduced into proteins of arbitrary size via mutagenesis and post-translational modification, this new probe could see wide use in characterizing the protein−lipid interactions of membrane proteins. American Chemical Society 2010-02-12 2010-03-04 /pmc/articles/PMC2836368/ /pubmed/20228945 http://dx.doi.org/10.1021/jz1000177 Text en Copyright © 2010 American Chemical Society http://pubs.acs.org This is an open-access article distributed under the ACS AuthorChoice Terms & Conditions. Any use of this article, must conform to the terms of that license which are available at http://pubs.acs.org.
spellingShingle McMahon, Heather A.
Alfieri, Katherine N.
Clark, Katherine A. A.
Londergan, Casey H.
Cyanylated Cysteine: A Covalently Attached Vibrational Probe of Protein−Lipid Contacts
title Cyanylated Cysteine: A Covalently Attached Vibrational Probe of Protein−Lipid Contacts
title_full Cyanylated Cysteine: A Covalently Attached Vibrational Probe of Protein−Lipid Contacts
title_fullStr Cyanylated Cysteine: A Covalently Attached Vibrational Probe of Protein−Lipid Contacts
title_full_unstemmed Cyanylated Cysteine: A Covalently Attached Vibrational Probe of Protein−Lipid Contacts
title_short Cyanylated Cysteine: A Covalently Attached Vibrational Probe of Protein−Lipid Contacts
title_sort cyanylated cysteine: a covalently attached vibrational probe of protein−lipid contacts
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2836368/
https://www.ncbi.nlm.nih.gov/pubmed/20228945
http://dx.doi.org/10.1021/jz1000177
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