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Use of MALDI-MS Combined with Differential Hydrogen–Deuterium Exchange for Semiautomated Protein Global Conformational Screening

Matrix-assisted laser desorption/ionization (MALDI) coupled with a time-of-flight (TOF) mass-spectrometry (MS) detector is acknowledged to be very useful for analysis of biological molecules. At the same time, hydrogen–deuterium exchange (HDX) is a well-known technique for studying protein higher-order structure. However, coupling MALDI with HDX has been challenging because of undesired back-exchange reactions during analysis. In this report, we survey an approach that utilizes MALDI coupled with an automated sample preparation to compare global conformational changes of proteins under different solution conditions using differential HDX. A nonaqueous matrix was proposed for MALDI sample preparation to minimize undesirable back-exchange. An automated experimental setup based on the use of a liquid-handling robot and automated data acquisition allowed for tracking protein conformational changes as a difference in the number of protons exchanged to deuterons at specified solution conditions. Experimental ti...

Authors:   Gregory F. Pirrone; Heather Wang; Nicole Canfield; Alexander S. Chin; Timothy A. Rhodes; Alexey A. Makarov
Journal:   Analytical Chemistry
Year:   2017
DOI:   10.1021/acs.analchem.7b01590
Publication date:   04-Aug-2017
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