Complete protein characterization using top-down mass spectrometry and ultraviolet photodissociation.

Complete protein characterization using top-down mass spectrometry and ultraviolet photodissociation.
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DOI:
10.1021/ja4029654
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发表时间:
2013-08-28
影响因子:
15
通讯作者:
Brodbelt, Jennifer S.
Brodbelt, Jennifer S.
中科院分区:
化学1区
文献类型:
--
作者:
Shaw, Jared B.;Li, Wenzong;Holden, Dustin D.;Zhang, Yan;Griep-Raming, Jens;Fellers, Ryan T.;Early, Bryan P.;Thomas, Paul M.;Kelleher, Neil L.;Brodbelt, Jennifer S.

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The top-down approach to proteomics offers compelling advantages due to the potential to provide complete characterization of protein sequence and post-translational modifications. Here we describe the implementation of 193 nm ultraviolet photodissociation (UVPD) in an Orbitrap mass spectrometer for characterization of intact proteins. Near-complete fragmentation of proteins up to 29 kDa is achieved with UVPD including the unambiguous localization of a single residue mutation and several protein modifications on Pin1 (Q13526), a protein implicated in the development of Alzheimer’s disease and in cancer pathogenesis. The 5 nanosecond, high-energy activation afforded by UVPD exhibits far less precursor ion-charge state dependence than conventional collision-based and electron-based dissociation methods.
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