title: External cavity-quantum cascade laser infrared spectroscopy for secondary structure analysis of proteins at low concentrations.
abstract: Fourier transform infrared (FTIR) and circular dichroism (CD) spectroscopy are analytical techniques employed for the analysis of protein secondary structure. The use of CD spectroscopy is limited to low protein concentrations (<2 mg ml(-1)), while FTIR spectroscopy is commonly used in a higher concentration range (>5 mg ml(-1)). Here we introduce a quantum cascade laser (QCL)-based IR transmission setup for analysis of protein and polypeptide secondary structure at concentrations as low as 0.25 mg ml(-1) in deuterated buffer solution. We present dynamic QCL-IR spectra of the temperature-induced α-helix to β-sheet transition of poly-L-lysine. The concentration dependence of the α-β transition temperature between 0.25 and 10 mg ml(-1) was investigated by QCL-IR, FTIR and CD spectroscopy. By using QCL-IR spectroscopy it is possible to perform IR spectroscopic analysis in the same concentration range as CD spectroscopy, thus enabling a combined analysis of biomolecules secondary structure by CD and IR spectroscopy.
authors.affiliation: Institute of Chemical Technologies and Analytics, Vienna University of Technology, Getreidemarkt 9/164-UPA, 1060 Vienna, Austria.
authors.forename: Andreas
authors.initials: A
authors.lastname: Schwaighofer
authors.affiliation: Institute of Chemical Technologies and Analytics, Vienna University of Technology, Getreidemarkt 9/164-UPA, 1060 Vienna, Austria.|Laboratorio de Desarrollo Analítico y Quimiometría, FBCB, Universidad Nacional del Litoral-CONICET, Ciudad Universitaria, 3000 Santa Fe, Argentina.
authors.forename: Mirta R
authors.initials: MR
authors.lastname: Alcaráz
authors.affiliation: Department of Chemistry, Institute of Biological Chemistry, University of Vienna, Währinger Straße 38, 1090 Vienna, Austria.
authors.forename: Can
authors.initials: C
authors.lastname: Araman
authors.affiliation: Laboratorio de Desarrollo Analítico y Quimiometría, FBCB, Universidad Nacional del Litoral-CONICET, Ciudad Universitaria, 3000 Santa Fe, Argentina.
authors.forename: Héctor
authors.initials: H
authors.lastname: Goicoechea
authors.affiliation: Institute of Chemical Technologies and Analytics, Vienna University of Technology, Getreidemarkt 9/164-UPA, 1060 Vienna, Austria.
authors.forename: Bernhard
authors.initials: B
authors.lastname: Lendl
mesh_terms: D002942:Circular Dichroism; D054023:Lasers, Semiconductor; D011107:Polylysine / Q000737:chemistry; D017433:Protein Structure, Secondary; D011506:Proteins / Q000737:chemistry*; D011789:Quantum Theory; D017550:Spectroscopy, Fourier Transform Infrared; D013696:Temperature
chemical_list: D011506:Proteins; D011107:Polylysine