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.jpg): Failed to open stream: No such file or directory in C:\Inetpub\vhosts\kidney.de\httpdocs\pget.php on line 117 J+Am+Soc+Mass+Spectrom
2016 ; 27
(3
): 462-73
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Ion Mobility Spectrometry-Hydrogen Deuterium Exchange Mass Spectrometry of
Anions: Part 3 Estimating Surface Area Exposure by Deuterium Uptake
#MMPMID26620531
J Am Soc Mass Spectrom
2016[Mar]; 27
(3
): 462-73
PMID26620531
show ga
Gas-phase hydrogen deuterium exchange (HDX), collision cross section (CCS)
measurement, and molecular dynamics simulation (MDS) techniques were utilized to
develop and compare three methods for estimating the relative surface area
exposure of separate peptide chains within bovine insulin ions. Electrosprayed [M
- 3H](3-) and [M - 5H](5-) insulin ions produced a single conformer type with
respective collision cross sections of 528 ± 5 Å(2) and 808 ± 2 Å(2). [M -
4H](4-) ions were comprised of more compact (? = 676 ± 3 Å(2)) and diffuse (i.e.,
more elongated, ? = 779 ± 3 Å(2)) ion conformer types. Ions were subjected to HDX
in the drift tube using D2O as the reagent gas. Collision-induced dissociation
was used to fragment mobility-selected, isotopically labeled [M - 4H](4-) and [M
- 5H](5-) ions into the protein subchains. Deuterium uptake levels of each chain
can be explained by limited inter-chain isotopic scrambling upon collisional
activation. Using nominal ion structures from MDS and a hydrogen accessibility
model, the deuterium uptake for each chain was correlated to its exposed surface
area. In separate experiments, the per-residue deuterium content for the
protonated and deprotonated ions of the synthetic peptide KKDDDDDIIKIIK were
compared. The differences in deuterium content indicated the regional HDX
accessibility for cations versus anions. Using ions of similar conformational
type, this comparison highlights the complementary nature of HDX data obtained
from positive- and negative-ion analysis.