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Minimizing Back Exchange in the Hydrogen Exchange-Mass Spectrometry Experiment

Overview of attention for article published in Journal of the American Society for Mass Spectrometry, September 2012
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Title
Minimizing Back Exchange in the Hydrogen Exchange-Mass Spectrometry Experiment
Published in
Journal of the American Society for Mass Spectrometry, September 2012
DOI 10.1007/s13361-012-0476-x
Pubmed ID
Authors

Benjamin T. Walters, Alec Ricciuti, Leland Mayne, S. Walter Englander

Abstract

The addition of mass spectrometry (MS) analysis to the hydrogen exchange (HX) proteolytic fragmentation experiment extends powerful HX methodology to the study of large biologically important proteins. A persistent problem is the degradation of HX information due to back exchange of deuterium label during the fragmentation-separation process needed to prepare samples for MS measurement. This paper reports a systematic analysis of the factors that influence back exchange (solution pH, ionic strength, desolvation temperature, LC column interaction, flow rates, system volume). The many peptides exhibit a range of back exchange due to intrinsic amino acid HX rate differences. Accordingly, large back exchange leads to large variability in D-recovery from one residue to another as well as one peptide to another that cannot be corrected for by reference to any single peptide-level measurement. The usual effort to limit back exchange by limiting LC time provides little gain. Shortening the LC elution gradient by 3-fold only reduced back exchange by ~2%, while sacrificing S/N and peptide count. An unexpected dependence of back exchange on ionic strength as well as pH suggests a strategy in which solution conditions are changed during sample preparation. Higher salt should be used in the first stage of sample preparation (proteolysis and trapping) and lower salt (<20 mM) and pH in the second stage before electrospray injection. Adjustment of these and other factors together with recent advances in peptide fragment detection yields hundreds of peptide fragments with D-label recovery of 90% ± 5%.

Twitter Demographics

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Mendeley readers

The data shown below were compiled from readership statistics for 108 Mendeley readers of this research output. Click here to see the associated Mendeley record.

Geographical breakdown

Country Count As %
United States 3 3%
Poland 1 <1%
United Kingdom 1 <1%
Denmark 1 <1%
India 1 <1%
Unknown 101 94%

Demographic breakdown

Readers by professional status Count As %
Student > Ph. D. Student 35 32%
Student > Master 23 21%
Researcher 17 16%
Unspecified 8 7%
Student > Doctoral Student 7 6%
Other 18 17%
Readers by discipline Count As %
Agricultural and Biological Sciences 36 33%
Chemistry 25 23%
Biochemistry, Genetics and Molecular Biology 23 21%
Unspecified 13 12%
Pharmacology, Toxicology and Pharmaceutical Science 5 5%
Other 6 6%

Attention Score in Context

This research output has an Altmetric Attention Score of 1. This is our high-level measure of the quality and quantity of online attention that it has received. This Attention Score, as well as the ranking and number of research outputs shown below, was calculated when the research output was last mentioned on 13 September 2012.
All research outputs
#3,657,071
of 4,512,164 outputs
Outputs from Journal of the American Society for Mass Spectrometry
#319
of 576 outputs
Outputs of similar age
#63,838
of 78,256 outputs
Outputs of similar age from Journal of the American Society for Mass Spectrometry
#46
of 54 outputs
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So far Altmetric has tracked 576 research outputs from this source. They receive a mean Attention Score of 1.3. This one is in the 1st percentile – i.e., 1% of its peers scored the same or lower than it.
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We're also able to compare this research output to 54 others from the same source and published within six weeks on either side of this one. This one is in the 1st percentile – i.e., 1% of its contemporaries scored the same or lower than it.