False EX1 signatures caused by sample carryover during HX MS analyses
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False EX1 signatures caused by sample carryover during HX MS analyses. / Fang, Jing; Rand, Kasper Dyrberg; Beuning, Penny J; Engen, John R.
In: International Journal of Mass Spectrometry, Vol. 302, No. 1-3, 2011, p. 19-25.Research output: Contribution to journal › Journal article › Research › peer-review
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TY - JOUR
T1 - False EX1 signatures caused by sample carryover during HX MS analyses
AU - Fang, Jing
AU - Rand, Kasper Dyrberg
AU - Beuning, Penny J
AU - Engen, John R
PY - 2011
Y1 - 2011
N2 - Discovery of EX1 kinetics in hydrogen exchange (HX) mass spectrometry (MS) experiments is rare. Proteins follow the EX1 kinetic regime when cooperative unfolding events simultaneously expose multiple residues to solvent such that they all become deuterated together before the region is able to refold. A number of factors can contribute to what we call "false EX1" in which it appears as though EX1 occurs in a protein when it probably does not. One of the contributors to false EX1 is peptide carryover between chromatographic runs. In this work, we explore the origins of peptide carryover in HX MS, describe how carryover causes mass spectra to indicate false EX1 kinetics and then describe an optimized washing protocol that can be used to eliminate peptide carryover. A series of solvent injections was developed and found to efficiently eliminate carryover signatures such that analysis of deuterium incorporation could be reliably followed for two proteins prone to high carryover.
AB - Discovery of EX1 kinetics in hydrogen exchange (HX) mass spectrometry (MS) experiments is rare. Proteins follow the EX1 kinetic regime when cooperative unfolding events simultaneously expose multiple residues to solvent such that they all become deuterated together before the region is able to refold. A number of factors can contribute to what we call "false EX1" in which it appears as though EX1 occurs in a protein when it probably does not. One of the contributors to false EX1 is peptide carryover between chromatographic runs. In this work, we explore the origins of peptide carryover in HX MS, describe how carryover causes mass spectra to indicate false EX1 kinetics and then describe an optimized washing protocol that can be used to eliminate peptide carryover. A series of solvent injections was developed and found to efficiently eliminate carryover signatures such that analysis of deuterium incorporation could be reliably followed for two proteins prone to high carryover.
U2 - 10.1016/j.ijms.2010.06.039
DO - 10.1016/j.ijms.2010.06.039
M3 - Journal article
C2 - 21643454
VL - 302
SP - 19
EP - 25
JO - International Journal of Mass Spectrometry
JF - International Journal of Mass Spectrometry
SN - 1387-3806
IS - 1-3
ER -
ID: 40129533