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Title

Modeling temperature-dependent protein structural transitions by combined near-IR and mid-IR spectroscopies and multivariate curve resolution

AuthorsNavea, Susana; Juan, Anna de; Tauler, Romà
KeywordsMIR
NIR
MCR−ALS
Protein structures
Multivariate Curve Resolution
Issue Date3-Sep-2003
PublisherAmerican Chemical Society
CitationAnalytical Chemistry 75(20): 5592-5601 (2003)
AbstractThe combination of near- and midinfrared spectroscopies (NIR and MIR) is proposed to monitor temperature-dependent transitions of proteins. These techniques offer a high discriminating power to distinguish among protein structural conformations but, in temperature-dependent processes, present the drawback associated with the intense and evolving absorption of the deuterium oxide, used as a solvent in the protein solutions. Multivariate curve resolution−alternating least squares (MCR−ALS) is chosen as the data analysis technique able to unravel the contributions of the pure protein and deuterium oxide species from the mixed raw experimental measurements. To do so, MCR−ALS works by analyzing simultaneously experiments from MIR and NIR on pure deuterium oxide solutions and protein solutions in D2O. This strategy has proven to be effective for modeling the protein process in the presence of D2O and, therefore, for avoiding the inclusion of artifacts in the data stemming from inadequate baseline corrections. The use of MIR and NIR and MCR−ALS has been tested in the study of the temperature-dependent evolution of β-lactoglobulin. Only the combined use of these two infrared techniques has allowed for the distinction of the three pure conformations involved in the process in the working thermal range: native, R-type state, and molten globule.
Description10 ppages.-- 8 figures.-- 1 table.-- PMID: 14710843 [PubMed].-- Printed version published Oct 15, 2003.
Publisher version (URL)http://dx.doi.org/10.1021/ac0343883
URIhttp://hdl.handle.net/10261/19381
DOI10.1021/ac0343883
ISSN0003-2700
E-ISSN1520-6882
Appears in Collections:(IDAEA) Artículos
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