Characterization of Native Protein Complexes Using Ultraviolet Photodissociation Mass Spectrometry

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• 作者：John P. O鈥橞rien ; Wenzong Li ; Yan Zhang ; Jennifer S. Brodbelt
• 刊名：Journal of the American Chemical Society
• 出版年：2014
• 出版时间：September 17, 2014
• 年：2014
• 卷：136
• 期：37
• 页码：12920-12928
• 全文大小：561K
• ISSN：1520-5126

文摘

Ultraviolet photodissociation (UVPD) mass spectrometry (MS) was used to characterize the sequences of proteins in native protein鈥搇igand and protein鈥損rotein complexes and to provide auxiliary information about the binding sites of the ligands and protein鈥損rotein interfaces. UVPD outperformed collisional induced dissociation (CID), higher-energy collisional dissociation (HCD), and electron transfer dissociation (ETD) in terms of yielding the most comprehensive diagnostic primary sequence information about the proteins in the complexes. UVPD also generated noncovalent fragment ions containing a portion of the protein still bound to the ligand which revealed some insight into the nature of the binding sites of myoglobin/heme, eIF4E/m⁷GTP, and human peptidyl-prolyl cis鈥搕rans isomerase 1 (Pin1) in complex with the peptide derived from the C-terminal domain of RNA polymerase II (CTD). Noncovalently bound protein鈥損rotein fragment ions from oligomeric 尾-lactoglobulin dimers and hexameric insulin complexes were also produced upon UVPD, providing some illumination of tertiary and quaternary protein structural features.