Modeling temperature-dependent protein structural transitions by combined near-IR and mid-IR spectroscopies and multivariate curve resolution
Modeling temperature-dependent protein structural transitions by combined near-IR and mid-IR spectroscopies and multivariate curve resolution
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DOI：
10.1021/ac0343883
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发表时间：
2003-10-15
影响因子：
7.4
通讯作者：
Tauler, R
中科院分区：
文献类型：
--
作者：
Navea, S;de Juan, A;Tauler, R
提出了近红外和中红外光谱（NIR和MIR）的组合来监测蛋白质的温度依赖性转变。这些技术提供了一个高的辨别能力，以区分蛋白质的结构构象，但在温度依赖性的过程中，目前的缺点与强烈的和不断变化的吸收的氧化氘，用作蛋白质溶液中的溶剂。多元曲线分辨率交替最小二乘法（MCR-ALS）被选为数据分析技术，能够解开的纯蛋白质和氧化氘物种的混合原始实验测量的贡献。为此，MCR-ALS通过同时分析来自MIR和NIR的纯氧化氘溶液和D2 O中的蛋白质溶液的实验来工作。这种策略已被证明是有效的蛋白质过程中的D2 O的存在下建模，因此，避免在数据中包含的工件源于不充分的基线校正。在β-乳球蛋白的温度依赖性演变研究中，已经测试了MIR和NIR以及MCR-ALS的使用。只有结合使用这两种红外技术，才能区分在工作温度范围内参与该过程的三种纯构象：天然的、R型状态和熔融球。
The 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 temperatare-dependent evolution of beta-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.