External-Cavity Quantum Cascade Laser Spectroscopy for Mid-IR Transmission Measurements of Proteins in Aqueous Solution

Autores
Alcaraz, Mirta Raquel; Schwaighofer, Andreas; Kristament, Christian; Ramer, Georg; Brandstetter, Markus; Goicoechea, Hector Casimiro; Lendl, Bernhard
Año de publicación
2015
Idioma
inglés
Tipo de recurso
artículo
Estado
versión publicada
Descripción
In this work, we report mid-IR transmissionmeasurements of the protein amide I band in aqueous solution atlarge optical paths. A tunable external-cavity quantum cascade laser(EC-QCL) operated in pulsed mode at room temperature allowedone to apply a path length of up to 38 μm, which is four times largerthan that applicable with conventional FT-IR spectrometers. Tominimize temperature-induced variations caused by backgroundabsorption of the ν2-vibration of water (HOH-bending) overlappingwith the amide I region, a highly stable temperature control unitwith relative temperature stability within 0.005 °C was developed.An advanced data processing protocol was established to overcomefluctuations in the fine structure of the emission curve that areinherent to the employed EC-QCL due to its mechanical instabilities. To allow for wavenumber accuracy, a spectral calibrationmethod has been elaborated to reference the acquired IR spectra to the absolute positions of the water vapor absorption bands.Employing this setup, characteristic spectral features of five well-studied proteins exhibiting different secondary structures couldbe measured at concentrations as low as 2.5 mg mL−1. This concentration range could previously only be accessed by IRmeasurements in D2O. Mathematical evaluation of the spectral overlap and comparison of second derivative spectra confirmexcellent agreement of the QCL transmission measurements with protein spectra acquired by FT-IR spectroscopy. This provesthe potential of the applied setup to monitor secondary structure changes of proteins in aqueous solution at extended opticalpath lengths, which allow experiments in flow through configuration.
Fil: Alcaraz, Mirta Raquel. Universidad Nacional del Litoral. Facultad de Bioquímica y Ciencias Biológicas. Departamento de Química. Cátedra de Química Analítica; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Santa Fe; Argentina
Fil: Schwaighofer, Andreas. Technische Universitat Wien; Austria
Fil: Kristament, Christian. Technische Universitat Wien; Austria
Fil: Ramer, Georg. Technische Universitat Wien; Austria
Fil: Brandstetter, Markus. Technische Universitat Wien; Austria
Fil: Goicoechea, Hector Casimiro. Universidad Nacional del Litoral. Facultad de Bioquímica y Ciencias Biológicas. Departamento de Química. Cátedra de Química Analítica; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Santa Fe; Argentina
Fil: Lendl, Bernhard. Technische Universitat Wien; Austria
Materia
Ec-Qcl
Mid-Ir
Infrared
Protein
Nivel de accesibilidad
acceso abierto
Condiciones de uso
https://creativecommons.org/licenses/by-nc-sa/2.5/ar/
Repositorio
CONICET Digital (CONICET)
Institución
Consejo Nacional de Investigaciones Científicas y Técnicas
OAI Identificador
oai:ri.conicet.gov.ar:11336/44058
Acceder
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oai_identifier_str oai:ri.conicet.gov.ar:11336/44058
network_acronym_str CONICETDig
repository_id_str 3498
network_name_str CONICET Digital (CONICET)
spelling External-Cavity Quantum Cascade Laser Spectroscopy for Mid-IR Transmission Measurements of Proteins in Aqueous SolutionAlcaraz, Mirta RaquelSchwaighofer, AndreasKristament, ChristianRamer, GeorgBrandstetter, MarkusGoicoechea, Hector CasimiroLendl, BernhardEc-QclMid-IrInfraredProteinhttps://purl.org/becyt/ford/1.4https://purl.org/becyt/ford/1In this work, we report mid-IR transmissionmeasurements of the protein amide I band in aqueous solution atlarge optical paths. A tunable external-cavity quantum cascade laser(EC-QCL) operated in pulsed mode at room temperature allowedone to apply a path length of up to 38 μm, which is four times largerthan that applicable with conventional FT-IR spectrometers. Tominimize temperature-induced variations caused by backgroundabsorption of the ν2-vibration of water (HOH-bending) overlappingwith the amide I region, a highly stable temperature control unitwith relative temperature stability within 0.005 °C was developed.An advanced data processing protocol was established to overcomefluctuations in the fine structure of the emission curve that areinherent to the employed EC-QCL due to its mechanical instabilities. To allow for wavenumber accuracy, a spectral calibrationmethod has been elaborated to reference the acquired IR spectra to the absolute positions of the water vapor absorption bands.Employing this setup, characteristic spectral features of five well-studied proteins exhibiting different secondary structures couldbe measured at concentrations as low as 2.5 mg mL−1. This concentration range could previously only be accessed by IRmeasurements in D2O. Mathematical evaluation of the spectral overlap and comparison of second derivative spectra confirmexcellent agreement of the QCL transmission measurements with protein spectra acquired by FT-IR spectroscopy. This provesthe potential of the applied setup to monitor secondary structure changes of proteins in aqueous solution at extended opticalpath lengths, which allow experiments in flow through configuration.Fil: Alcaraz, Mirta Raquel. Universidad Nacional del Litoral. Facultad de Bioquímica y Ciencias Biológicas. Departamento de Química. Cátedra de Química Analítica; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Santa Fe; ArgentinaFil: Schwaighofer, Andreas. Technische Universitat Wien; AustriaFil: Kristament, Christian. Technische Universitat Wien; AustriaFil: Ramer, Georg. Technische Universitat Wien; AustriaFil: Brandstetter, Markus. Technische Universitat Wien; AustriaFil: Goicoechea, Hector Casimiro. Universidad Nacional del Litoral. Facultad de Bioquímica y Ciencias Biológicas. Departamento de Química. Cátedra de Química Analítica; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Santa Fe; ArgentinaFil: Lendl, Bernhard. Technische Universitat Wien; AustriaAmerican Chemical Society2015-06info:eu-repo/semantics/articleinfo:eu-repo/semantics/publishedVersionhttp://purl.org/coar/resource_type/c_6501info:ar-repo/semantics/articuloapplication/pdfapplication/pdfhttp://hdl.handle.net/11336/44058Alcaraz, Mirta Raquel; Schwaighofer, Andreas; Kristament, Christian; Ramer, Georg; Brandstetter, Markus; et al.; External-Cavity Quantum Cascade Laser Spectroscopy for Mid-IR Transmission Measurements of Proteins in Aqueous Solution; American Chemical Society; Analytical Chemistry; 87; 13; 6-2015; 6980-69870003-2700CONICET DigitalCONICETenginfo:eu-repo/semantics/altIdentifier/doi/10.1021/acs.analchem.5b01738info:eu-repo/semantics/altIdentifier/url/https://pubs.acs.org/doi/10.1021/acs.analchem.5b01738info:eu-repo/semantics/openAccesshttps://creativecommons.org/licenses/by-nc-sa/2.5/ar/reponame:CONICET Digital (CONICET)instname:Consejo Nacional de Investigaciones Científicas y Técnicas2025-09-03T10:02:11Zoai:ri.conicet.gov.ar:11336/44058instacron:CONICETInstitucionalhttp://ri.conicet.gov.ar/Organismo científico-tecnológicoNo correspondehttp://ri.conicet.gov.ar/oai/requestdasensio@conicet.gov.ar; lcarlino@conicet.gov.arArgentinaNo correspondeNo correspondeNo correspondeopendoar:34982025-09-03 10:02:12.159CONICET Digital (CONICET) - Consejo Nacional de Investigaciones Científicas y Técnicasfalse
dc.title.none.fl_str_mv External-Cavity Quantum Cascade Laser Spectroscopy for Mid-IR Transmission Measurements of Proteins in Aqueous Solution
title External-Cavity Quantum Cascade Laser Spectroscopy for Mid-IR Transmission Measurements of Proteins in Aqueous Solution
spellingShingle External-Cavity Quantum Cascade Laser Spectroscopy for Mid-IR Transmission Measurements of Proteins in Aqueous Solution
Alcaraz, Mirta Raquel
Ec-Qcl
Mid-Ir
Infrared
Protein
title_short External-Cavity Quantum Cascade Laser Spectroscopy for Mid-IR Transmission Measurements of Proteins in Aqueous Solution
title_full External-Cavity Quantum Cascade Laser Spectroscopy for Mid-IR Transmission Measurements of Proteins in Aqueous Solution
title_fullStr External-Cavity Quantum Cascade Laser Spectroscopy for Mid-IR Transmission Measurements of Proteins in Aqueous Solution
title_full_unstemmed External-Cavity Quantum Cascade Laser Spectroscopy for Mid-IR Transmission Measurements of Proteins in Aqueous Solution
title_sort External-Cavity Quantum Cascade Laser Spectroscopy for Mid-IR Transmission Measurements of Proteins in Aqueous Solution
dc.creator.none.fl_str_mv Alcaraz, Mirta Raquel
Schwaighofer, Andreas
Kristament, Christian
Ramer, Georg
Brandstetter, Markus
Goicoechea, Hector Casimiro
Lendl, Bernhard
author Alcaraz, Mirta Raquel
author_facet Alcaraz, Mirta Raquel
Schwaighofer, Andreas
Kristament, Christian
Ramer, Georg
Brandstetter, Markus
Goicoechea, Hector Casimiro
Lendl, Bernhard
author_role author
author2 Schwaighofer, Andreas
Kristament, Christian
Ramer, Georg
Brandstetter, Markus
Goicoechea, Hector Casimiro
Lendl, Bernhard
author2_role author
author
author
author
author
author
dc.subject.none.fl_str_mv Ec-Qcl
Mid-Ir
Infrared
Protein
topic Ec-Qcl
Mid-Ir
Infrared
Protein
purl_subject.fl_str_mv https://purl.org/becyt/ford/1.4
https://purl.org/becyt/ford/1
dc.description.none.fl_txt_mv In this work, we report mid-IR transmissionmeasurements of the protein amide I band in aqueous solution atlarge optical paths. A tunable external-cavity quantum cascade laser(EC-QCL) operated in pulsed mode at room temperature allowedone to apply a path length of up to 38 μm, which is four times largerthan that applicable with conventional FT-IR spectrometers. Tominimize temperature-induced variations caused by backgroundabsorption of the ν2-vibration of water (HOH-bending) overlappingwith the amide I region, a highly stable temperature control unitwith relative temperature stability within 0.005 °C was developed.An advanced data processing protocol was established to overcomefluctuations in the fine structure of the emission curve that areinherent to the employed EC-QCL due to its mechanical instabilities. To allow for wavenumber accuracy, a spectral calibrationmethod has been elaborated to reference the acquired IR spectra to the absolute positions of the water vapor absorption bands.Employing this setup, characteristic spectral features of five well-studied proteins exhibiting different secondary structures couldbe measured at concentrations as low as 2.5 mg mL−1. This concentration range could previously only be accessed by IRmeasurements in D2O. Mathematical evaluation of the spectral overlap and comparison of second derivative spectra confirmexcellent agreement of the QCL transmission measurements with protein spectra acquired by FT-IR spectroscopy. This provesthe potential of the applied setup to monitor secondary structure changes of proteins in aqueous solution at extended opticalpath lengths, which allow experiments in flow through configuration.
Fil: Alcaraz, Mirta Raquel. Universidad Nacional del Litoral. Facultad de Bioquímica y Ciencias Biológicas. Departamento de Química. Cátedra de Química Analítica; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Santa Fe; Argentina
Fil: Schwaighofer, Andreas. Technische Universitat Wien; Austria
Fil: Kristament, Christian. Technische Universitat Wien; Austria
Fil: Ramer, Georg. Technische Universitat Wien; Austria
Fil: Brandstetter, Markus. Technische Universitat Wien; Austria
Fil: Goicoechea, Hector Casimiro. Universidad Nacional del Litoral. Facultad de Bioquímica y Ciencias Biológicas. Departamento de Química. Cátedra de Química Analítica; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Santa Fe; Argentina
Fil: Lendl, Bernhard. Technische Universitat Wien; Austria
description In this work, we report mid-IR transmissionmeasurements of the protein amide I band in aqueous solution atlarge optical paths. A tunable external-cavity quantum cascade laser(EC-QCL) operated in pulsed mode at room temperature allowedone to apply a path length of up to 38 μm, which is four times largerthan that applicable with conventional FT-IR spectrometers. Tominimize temperature-induced variations caused by backgroundabsorption of the ν2-vibration of water (HOH-bending) overlappingwith the amide I region, a highly stable temperature control unitwith relative temperature stability within 0.005 °C was developed.An advanced data processing protocol was established to overcomefluctuations in the fine structure of the emission curve that areinherent to the employed EC-QCL due to its mechanical instabilities. To allow for wavenumber accuracy, a spectral calibrationmethod has been elaborated to reference the acquired IR spectra to the absolute positions of the water vapor absorption bands.Employing this setup, characteristic spectral features of five well-studied proteins exhibiting different secondary structures couldbe measured at concentrations as low as 2.5 mg mL−1. This concentration range could previously only be accessed by IRmeasurements in D2O. Mathematical evaluation of the spectral overlap and comparison of second derivative spectra confirmexcellent agreement of the QCL transmission measurements with protein spectra acquired by FT-IR spectroscopy. This provesthe potential of the applied setup to monitor secondary structure changes of proteins in aqueous solution at extended opticalpath lengths, which allow experiments in flow through configuration.
publishDate 2015
dc.date.none.fl_str_mv 2015-06
dc.type.none.fl_str_mv info:eu-repo/semantics/article
info:eu-repo/semantics/publishedVersion
http://purl.org/coar/resource_type/c_6501
info:ar-repo/semantics/articulo
format article
status_str publishedVersion
dc.identifier.none.fl_str_mv http://hdl.handle.net/11336/44058
Alcaraz, Mirta Raquel; Schwaighofer, Andreas; Kristament, Christian; Ramer, Georg; Brandstetter, Markus; et al.; External-Cavity Quantum Cascade Laser Spectroscopy for Mid-IR Transmission Measurements of Proteins in Aqueous Solution; American Chemical Society; Analytical Chemistry; 87; 13; 6-2015; 6980-6987
0003-2700
CONICET Digital
CONICET
url http://hdl.handle.net/11336/44058
identifier_str_mv Alcaraz, Mirta Raquel; Schwaighofer, Andreas; Kristament, Christian; Ramer, Georg; Brandstetter, Markus; et al.; External-Cavity Quantum Cascade Laser Spectroscopy for Mid-IR Transmission Measurements of Proteins in Aqueous Solution; American Chemical Society; Analytical Chemistry; 87; 13; 6-2015; 6980-6987
0003-2700
CONICET Digital
CONICET
dc.language.none.fl_str_mv eng
language eng
dc.relation.none.fl_str_mv info:eu-repo/semantics/altIdentifier/doi/10.1021/acs.analchem.5b01738
info:eu-repo/semantics/altIdentifier/url/https://pubs.acs.org/doi/10.1021/acs.analchem.5b01738
dc.rights.none.fl_str_mv info:eu-repo/semantics/openAccess
https://creativecommons.org/licenses/by-nc-sa/2.5/ar/
eu_rights_str_mv openAccess
rights_invalid_str_mv https://creativecommons.org/licenses/by-nc-sa/2.5/ar/
dc.format.none.fl_str_mv application/pdf
application/pdf
dc.publisher.none.fl_str_mv American Chemical Society
publisher.none.fl_str_mv American Chemical Society
dc.source.none.fl_str_mv reponame:CONICET Digital (CONICET)
instname:Consejo Nacional de Investigaciones Científicas y Técnicas
reponame_str CONICET Digital (CONICET)
collection CONICET Digital (CONICET)
instname_str Consejo Nacional de Investigaciones Científicas y Técnicas
repository.name.fl_str_mv CONICET Digital (CONICET) - Consejo Nacional de Investigaciones Científicas y Técnicas
repository.mail.fl_str_mv dasensio@conicet.gov.ar; lcarlino@conicet.gov.ar
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score 13.13397

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