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Amide hydrogen exchange shows that malate dehydrogenase is a folded monomer at pH 5
Author(s) -
Chen Jiwen,
Smith David L.
Publication year - 2001
Publication title -
protein science
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 3.353
H-Index - 175
eISSN - 1469-896X
pISSN - 0961-8368
DOI - 10.1110/ps.53201
Subject(s) - malate dehydrogenase , chemistry , dimer , hydrogen–deuterium exchange , monomer , groel , solvent , chaperonin , hydrogen , crystallography , biochemistry , enzyme , organic chemistry , protein folding , polymer , escherichia coli , gene
Although there is general agreement that native mitochondrial malate dehydrogenase (MDH) exists as a dimer at pH 7, its aggregation state at pH 5 is less certain. The present amide hydrogen exchange study was performed to determine whether MDH remains a dimer at pH 5. To detect pH‐induced changes in solvent accessibility, MDH was exposed to D 2 O at pH 5 or 7, then fragmented with pepsin into peptides that were analyzed by mass spectrometry. Even after adjustments for the effect of pH on the intrinsic rate of hydrogen exchange, large increases in deuterium levels were found at pH 5 only in peptic fragments derived from the subunit binding surface of MDH. In parallel experiments, elevated deuterium levels were also found in the same regions of MDH monomer trapped inside a mutant form of the chaperonin GroEL. This selective increase in hydrogen exchange rates, which was attributed to increased solvent accessibility of these regions, provides new evidence that MDH is a monomer at pH 5.