Complete Protein Characterization Using Top-Down Mass Spectrometry and Ultraviolet Photodissociation

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• 作者：Jared B. Shaw ; Wenzong Li ; Dustin D. Holden ; Yan Zhang ; Jens Griep-Raming ; Ryan T. Fellers ; Bryan P. Early ; Paul M. Thomas ; Neil L. Kelleher ; Jennifer S. Brodbelt
• 刊名：Journal of the American Chemical Society
• 出版年：2013
• 出版时间：August 28, 2013
• 年：2013
• 卷：135
• 期：34
• 页码：12646-12651
• 全文大小：425K
• 年卷期：v.135,no.34(August 28, 2013)
• ISSN：1520-5126

文摘

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鈥檚 disease and in cancer pathogenesis. The 5 ns, high-energy activation afforded by UVPD exhibits far less precursor ion-charge state dependence than conventional collision- and electron-based dissociation methods.