First-Principles Collision Cross Section Measurements of Large Proteins and Protein Complexes

Jacob W. Mccabe, Christopher S. Mallis, Klaudia I. Kocurek, Michael L. Poltash, Mehdi Shirzadeh, Michael J. Hebert, Liqi Fan, Thomas E. Walker, Xueyun Zheng, Ting Jiang, Shiyu Dong, Cheng Wei Lin, Arthur Laganowsky, David H. Russell*

*此作品的通讯作者

科研成果: 期刊稿件文章同行评审

25 引用 (Scopus)

摘要

Rotationally averaged collision cross section (CCS) values for a series of proteins and protein complexes ranging in size from 8.6 to 810 kDa are reported. The CCSs were obtained using a native electrospray ionization drift tube ion mobility-Orbitrap mass spectrometer specifically designed to enhance sensitivity while having high-resolution ion mobility and mass capabilities. Periodic focusing (PF)-drift tube (DT)-ion mobility (IM) provides first-principles determination of the CCS of large biomolecules that can then be used as CCS calibrants. The experimental, first-principles CCS values are compared to previously reported experimentally determined and computationally calculated CCS using projected superposition approximation (PSA), the Ion Mobility Projection Approximation Calculation Tool (IMPACT), and Collidoscope. Experimental CCS values are generally in agreement with previously reported CCSs, with values falling within ∼5.5%. In addition, an ion mobility resolution (CCS centroid divided by CCS fwhm) of ∼60 is obtained for pyruvate kinase (MW ∼233 kDa); however, ion mobility resolution for bovine serum albumin (MW ∼68 kDa) is less than ∼20, which arises from sample impurities and underscores the importance of sample quality. The high resolution afforded by the ion mobility-Orbitrap mass analyzer provides new opportunities to understand the intricate details of protein complexes such as the impact of post-translational modifications (PTMs), stoichiometry, and conformational changes induced by ligand binding.

源语言英语
页(从-至)11155-11163
页数9
期刊Analytical Chemistry
92
16
DOI
出版状态已出版 - 18 8月 2020
已对外发布

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