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Elucidating Escherichia coli Proteoform Families Using Intact-Mass Proteomics and a Global PTM Discovery Database
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posted on 2017-10-02, 00:00 authored by Yunxiang Dai, Michael R. Shortreed, Mark Scalf, Brian L. Frey, Anthony J. Cesnik, Stefan Solntsev, Leah V. Schaffer, Lloyd M. SmithA proteoform family is a group of
related molecular forms of a
protein (proteoforms) derived from the same gene. We have previously
described a strategy to identify proteoforms and elucidate proteoform
families in complex mixtures of intact proteins. The strategy is based
upon measurements of two properties for each proteoform: (i) the accurate
proteoform intact-mass, measured by liquid chromatography/mass spectrometry
(LC–MS), and (ii) the number of lysine residues in each proteoform,
determined using an isotopic labeling approach. These measured properties
are then compared with those extracted from a catalog of theoretical
proteoforms containing protein sequences and localized post-translational
modifications (PTMs) for the organism under study. A match between
the measured properties and those in the catalog constitutes an identification
of the proteoform. In the present study, this strategy is extended
by utilizing a global PTM discovery database and is applied to the
widely studied model organism Escherichia coli, providing
the most comprehensive elucidation of E. coli proteoforms
and proteoform families to date.
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Elucidating Escherichia coli Proteoform Familiesmodel organism Escherichia colilysine residuesproteoform familiesprotein sequencescoli proteoformsIntact-Mass ProteomicsPTM discovery databaseLCpost-translational modificationsproteoform intact-massproteoform familyGlobal PTM Discovery Databaseelucidate proteoform families
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