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Improved Sequence Analysis of Intact Proteins by Parallel Ion Parking during Electron Transfer Dissociation.


ABSTRACT: Electron transfer dissociation (ETD) is an analytically useful tool for primary structure interrogation of intact proteins, but its utility is limited by higher-order reactions with the products. To inhibit these higher-order reactions, first-generation fragment ions are kinetically excited by applying an experimentally tailored parallel ion parking waveform during ETD (ETD-PIP). In combination with subsequent ion/ion proton transfer reactions, precursor-to-product conversion was maximized as evidenced by the consumption of more than 90% of the 21 kDa Protein G precursor to form ETD product ions. The employment of ETD-PIP increased sequence coverage to 90% from 80% with standard ETD. Additionally, the inhibition of sequential electron transfers was reflected in the high number of complementary ion pairs from ETD-PIP (90%) compared to standard ETD (39%).

SUBMITTER: Duselis EM 

PROVIDER: S-EPMC8855838 | biostudies-literature | 2021 Nov

REPOSITORIES: biostudies-literature

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Improved Sequence Analysis of Intact Proteins by Parallel Ion Parking during Electron Transfer Dissociation.

Duselis Elizabeth M EM   Panepinto Maria C MC   Syka John E P JEP   Mullen Christopher C   D'Ippolito Robert A RA   English A Michelle AM   Ugrin Scott A SA   Shabanowitz Jeffrey J   Hunt Donald F DF  

Analytical chemistry 20211117 47


Electron transfer dissociation (ETD) is an analytically useful tool for primary structure interrogation of intact proteins, but its utility is limited by higher-order reactions with the products. To inhibit these higher-order reactions, first-generation fragment ions are kinetically excited by applying an experimentally tailored parallel ion parking waveform during ETD (ETD-PIP). In combination with subsequent ion/ion proton transfer reactions, precursor-to-product conversion was maximized as ev  ...[more]

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