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首页> 美国卫生研究院文献>other >An On-Line High-Pressure Digestion System for Protein Characterization by Hydrogen/Deuterium Exchange and Mass Spectrometry
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An On-Line High-Pressure Digestion System for Protein Characterization by Hydrogen/Deuterium Exchange and Mass Spectrometry

机译：用于氢气/氘交换和质谱法的蛋白质表征的在线高压消化系统

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The rapid and complete digestion of proteins is important when protein characterization by hydrogen deuterium exchange (HDX) is coupled with mass spectrometry. We developed a single-pump, on-line, high pressure digestion system that relies on UPLC technology to aid in the digestion of proteins. Two model proteins, Aβ1–42 and NBSA, were used to demonstrate the efficacy of the high pressure system. Both model proteins readily aggregate and are difficult to digest under normal conditions. Our high pressure system successfully digests these proteins into small, overlapping peptides. The extra information afforded by overlapping peptides allows us to pinpoint HDX protection to protein segments smaller than the digested peptide. The calculated average segment length (ASL) for both model proteins decreased by 2-fold for high pressure digestion compared to digestion at ambient pressure.

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