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Studying protein‐carbohydrate interactions by amide hydrogen/deuterium exchange mass spectrometry
Author(s) -
King Daniel,
Lumpkin Michelle,
Bergmann Carl,
Orlando Ron
Publication year - 2002
Publication title -
rapid communications in mass spectrometry
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.528
H-Index - 136
eISSN - 1097-0231
pISSN - 0951-4198
DOI - 10.1002/rcm.746
Subject(s) - chemistry , hydrogen–deuterium exchange , amide , deuterium , substrate (aquarium) , carbohydrate , mass spectrometry , stereochemistry , chromatography , organic chemistry , physics , oceanography , quantum mechanics , geology
Abstract Protein‐carbohydrate interactions play a significant role in biological processes. Presented here is the novel application of amide hydrogen/deuterium exchange mass spectrometry (amide exchange‐MS) to the study of the interaction between a protein and its carbohydrate substrate. The degree of deuterium incorporation into hen egg lysozyme was monitored with and without substrate to verify that a carbohydrate can provide sufficiently stable protection of the amide hydrogen atoms in a protein's backbone from exchange with deuterated solvent. The substrate protected a number of amide hydrogens from exchange, implying that protein‐carbohydrate binding systems will be compatible with amide exchange‐MS. Endopolygalacturonase‐II (EPG‐II) from Aspergillus niger , a pectin‐degrading enzyme, was chosen as the first carbohydrate‐binding system to be extensively studied using quenched amide exchange‐MS. Monitoring the changes in deuterium incorporation of EPG‐II in the presence and absence of an oligomer of galacturonic acid implied the location of substrate binding. This study demonstrates the ability of amide exchange‐MS to investigate protein‐carbohydrate interactions. Copyright © 2002 John Wiley & Sons, Ltd.