دورية أكاديمية
Enhanced Declustering Enables Native Top-Down Analysis of Membrane Protein Complexes using Ion-Mobility Time-Aligned Fragmentation.
| العنوان: | Enhanced Declustering Enables Native Top-Down Analysis of Membrane Protein Complexes using Ion-Mobility Time-Aligned Fragmentation. |
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| المؤلفون: | Sokratous K; OMass Therapeutics, Chancellor Court, John Smith Drive, ARC Oxford OX4 2GX, United Kingdom., Cooper-Shepherd DA; Waters Corporation, Stamford Avenue, Altrincham Road, Wilmslow SK9 4AX, United Kingdom., Ujma J; Waters Corporation, Stamford Avenue, Altrincham Road, Wilmslow SK9 4AX, United Kingdom., Qu F; OMass Therapeutics, Chancellor Court, John Smith Drive, ARC Oxford OX4 2GX, United Kingdom., Giles K; Waters Corporation, Stamford Avenue, Altrincham Road, Wilmslow SK9 4AX, United Kingdom., Ben-Younis A; OMass Therapeutics, Chancellor Court, John Smith Drive, ARC Oxford OX4 2GX, United Kingdom., Hensen M; OMass Therapeutics, Chancellor Court, John Smith Drive, ARC Oxford OX4 2GX, United Kingdom., Langridge JI; Waters Corporation, Stamford Avenue, Altrincham Road, Wilmslow SK9 4AX, United Kingdom., Gault J; OMass Therapeutics, Chancellor Court, John Smith Drive, ARC Oxford OX4 2GX, United Kingdom., Jazayeri A; OMass Therapeutics, Chancellor Court, John Smith Drive, ARC Oxford OX4 2GX, United Kingdom., Liko I; OMass Therapeutics, Chancellor Court, John Smith Drive, ARC Oxford OX4 2GX, United Kingdom., Hopper JTS; OMass Therapeutics, Chancellor Court, John Smith Drive, ARC Oxford OX4 2GX, United Kingdom. |
| المصدر: | Journal of the American Society for Mass Spectrometry [J Am Soc Mass Spectrom] 2024 Aug 07; Vol. 35 (8), pp. 1891-1901. Date of Electronic Publication: 2024 Jul 15. |
| نوع المنشور: | Journal Article |
| اللغة: | English |
| بيانات الدورية: | Publisher: ACS Publications Country of Publication: United States NLM ID: 9010412 Publication Model: Print-Electronic Cited Medium: Internet ISSN: 1879-1123 (Electronic) Linking ISSN: 10440305 NLM ISO Abbreviation: J Am Soc Mass Spectrom Subsets: MEDLINE |
| أسماء مطبوعة: | Publication: 2020- : Washington, DC : ACS Publications Original Publication: New York, NY : Elsevier, c1990- |
| مواضيع طبية MeSH: | Membrane Proteins*/chemistry , Membrane Proteins*/analysis , Ion Mobility Spectrometry*/methods, Micelles ; Mass Spectrometry/methods ; Detergents/chemistry ; Ions/chemistry |
| مستخلص: | Native mass spectrometry (MS) is proving to be a disruptive technique for studying the interactions of proteins, necessary for understanding the functional roles of these biomolecules. Recent research is expanding the application of native MS towards membrane proteins directly from isolated membrane preparations or from purified detergent micelles. The former results in complex spectra comprising several heterogeneous protein complexes; the latter enables therapeutic protein targets to be screened against multiplexed preparations of compound libraries. In both cases, the resulting spectra are increasingly complex to assign/interpret, and the key to these new directions of native MS research is the ability to perform native top-down analysis, which allows unambiguous peak assignment. To achieve this, detergent removal is necessary prior to MS analyzers, which allow selection of specific m / z values, representing the parent ion for downstream activation. Here, we describe a novel, enhanced declustering (ED) device installed into the first pumping region of a cyclic IMS-enabled mass spectrometry platform. The device enables declustering of ions prior to the quadrupole by imparting collisional activation through an oscillating electric field applied between two parallel plates. The positioning of the device enables liberation of membrane protein ions from detergent micelles. Quadrupole selection can now be utilized to isolate protein-ligand complexes, and downstream collision cells enable the dissociation and identification of binding partners. We demonstrate that ion mobility (IM) significantly aids in the assignment of top-down spectra, aligning fragments to their corresponding parent ions by means of IM drift time. Using this approach, we were able to confidently assign and identify a novel hit compound against Pf MATE, obtained from multiplexed ligand libraries. |
| فهرسة مساهمة: | Keywords: ion-mobility spectrometry; membrane protein; native mass spectrometry; top down |
| المشرفين على المادة: | 0 (Membrane Proteins) 0 (Micelles) 0 (Detergents) 0 (Ions) |
| تواريخ الأحداث: | Date Created: 20240715 Date Completed: 20240807 Latest Revision: 20240807 |
| رمز التحديث: | 20240807 |
| DOI: | 10.1021/jasms.4c00190 |
| PMID: | 39007842 |
| قاعدة البيانات: | MEDLINE |
Find this article in full text from Springer Verlag. 
Find this issue from the American Chemical Society | تدمد: | 1879-1123 |
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| DOI: | 10.1021/jasms.4c00190 |