Thermal Unfolding of Proteins Studied by Coupled Reversed-Phase HPLC-Electrospray Ionization Mass Spectrometry Techniques Based on Isotope Exchange Effects

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• 作者：Milton T. W. Hearn ; Joselito P. Quirino ; James Whisstock ; and Shigeru Terabe
• 刊名：Analytical Chemistry
• 出版年：2002
• 出版时间：March 15, 2002
• 年：2002
• 卷：74
• 期：6
• 页码：1467 - 1475
• 全文大小：103K
• 年卷期：v.74,no.6(March 15, 2002)
• ISSN：1520-6882

文摘

In this study, a deuterium exchange procedure has beenemployed to evaluate the thermal stability of globularproteins under conditions that replicate their interactivebehavior in reversed-phase high performance chromatographic (RP-HPLC) systems. In particular, this investigation has permitted the conformational stability of twoproteins, hen egg white lysozyme (HEWL) and horse heartmyoglobin (HMYO) to be examined under different temperature and low-pH solvent regimes. The results confirmthat this experimental approach provides an efficientstrategy to explore fundamental conformational featuresof polypeptides or proteins in their folded and partialunfolded states under these interactive conditions. Inparticular, this analytical procedure permits insight to bereadily gained into the processes that occur when polypeptides and globular proteins interact with lipophilic liquid/solid interfaces in the presence of water-organic solventmixtures at different temperatures.