Titelaufnahme

Titel
EC-QCL mid-IR transmission spectroscopy for monitoring dynamic changes of protein secondary structure in aqueous solution on the example of -aggregation in alcohol-denaturated -chymotrypsin
VerfasserLendl, Bernhard In der Gemeinsamen Normdatei der DNB nachschlagen ; Alcaráz, Mirta R. ; Schwaighofer, Andreas ; Goicoechea, Héctor
Erschienen in
Analytical and Bioanalytical Chemistry, Berlin ; Heidelberg 2016, Jg. 408, H. 15, S. 3933-3941
ErschienenSpringer 2016
Ausgabe
Published version
SpracheEnglisch
DokumenttypAufsatz in einer Zeitschrift
Schlagwörter (EN)Quantum cascade laser / Infrared spectroscopy / Multivariate curve resolution-alternating / least squares / Protein secondary structure / Aggregation / ,2,2-Trifluoroethanol
ISSN1618-2650
URNurn:nbn:at:at-ubtuw:3-1668 Persistent Identifier (URN)
DOIdoi:10.1007/s00216-016-9464-5 
Lizenz
CC-BY-Lizenz (4.0)Creative Commons Namensnennung 4.0 International Lizenz
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 Das Werk ist frei verfügbar
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EC-QCL mid-IR transmission spectroscopy for monitoring dynamic changes of protein secondary structure in aqueous solution on the example of -aggregation in alcohol-denaturated -chymotrypsin [0.9 mb]
Supplementary material [0.16 mb]
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Zusammenfassung (Englisch)

In this work, a novel EC-QCL-based setup for mid-IR transmission measurements in the amide I region is introduced for monitoring dynamic changes in secondary structure of proteins. For this purpose, -chymotrypsin (aCT) acts as a model protein, which gradually forms intermolecular -sheet aggregates after adopting a non-native -helical structure induced by exposure to 50 % TFE. In order to showcase the versatility of the presented setup, the effects of varying pH values and protein concentration on the rate of -aggregation were studied. The influence of the pH value on the initial reaction rate was studied in the range of pH 5.88.2. Results indicate an increased aggregation rate at elevated pH values. Furthermore, the widely accessible concentration range of the laser-based IR transmission setup was utilized to investigate -aggregation across a concentration range of 560 mg mL1. For concentrations lower than 20 mg mL1, the aggregation rate appears to be independent of concentration. At higher values, the reaction rate increases linearly with protein concentration. Extended MCR-ALS was employed to obtain pure spectral and concentration profiles of the temporal transition between -helices and intermolecular -sheets. Comparison of the global solutions obtained by the modelled data with results acquired by the laser-based IR transmission setup at different conditions shows excellent agreement. This demonstrates the potential and versatility of the EC-QCL-based IR transmission setup to monitor dynamic changes of protein secondary structure in aqueous solution at varying conditions and across a wide concentration range.

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