التفاصيل البيبلوغرافية
| العنوان: |
Expanding the Depth and Sensitivity of Cross-Link Identification by Differential Ion Mobility Using High-Field Asymmetric Waveform Ion Mobility Spectrometry. |
| المؤلفون: |
Schnirch L; Department of Chemical Biology, Leibniz-Forschungsinstitut für Molekulare Pharmakologie (FMP), Robert-Roessle-Str. 10, 13125 Berlin, Germany., Nadler-Holly M; Department of Chemical Biology, Leibniz-Forschungsinstitut für Molekulare Pharmakologie (FMP), Robert-Roessle-Str. 10, 13125 Berlin, Germany., Siao SW; Department of Chemical Biology, Leibniz-Forschungsinstitut für Molekulare Pharmakologie (FMP), Robert-Roessle-Str. 10, 13125 Berlin, Germany.; Max Planck Unit for the Science of Pathogens, Charitéplatz 1, 10117 Berlin, Germany., Frese CK; Max Planck Unit for the Science of Pathogens, Charitéplatz 1, 10117 Berlin, Germany., Viner R; Thermo Fisher Scientific, 355 River Oaks Pkwy., San Jose, California 95134, United States., Liu F; Department of Chemical Biology, Leibniz-Forschungsinstitut für Molekulare Pharmakologie (FMP), Robert-Roessle-Str. 10, 13125 Berlin, Germany. |
| المصدر: |
Analytical chemistry [Anal Chem] 2020 Aug 04; Vol. 92 (15), pp. 10495-10503. Date of Electronic Publication: 2020 Jul 23. |
| نوع المنشور: |
Journal Article; Research Support, Non-U.S. Gov't |
| اللغة: |
English |
| بيانات الدورية: |
Publisher: American Chemical Society Country of Publication: United States NLM ID: 0370536 Publication Model: Print-Electronic Cited Medium: Internet ISSN: 1520-6882 (Electronic) Linking ISSN: 00032700 NLM ISO Abbreviation: Anal Chem Subsets: MEDLINE |
| أسماء مطبوعة: |
Original Publication: Washington, American Chemical Society. |
| مواضيع طبية MeSH: |
Ion Mobility Spectrometry/*methods , Peptides/*chemistry, Chemical Fractionation ; Chromatography/methods ; HEK293 Cells ; Humans |
| مستخلص: |
In cross-linking mass spectrometry (XL-MS), the depth and sensitivity of cross-link detection is often limited by the low abundance of cross-links compared to non-cross-linked peptides in the digestion mixture. To improve the identification efficiency of cross-links, here, we present a gas-phase separation strategy using high-field asymmetric waveform ion mobility spectrometry (FAIMS) coupled to the Orbitrap Tribrid mass spectrometers. By enabling an additional peptide separation step in the gas phase using the FAIMS device, we increase the number of cross-link identifications by 22% for a medium complex sample and 59% for strong cation exchange-fractionated HEK293T cell lysate in XL-MS experiments using disuccinimidyl sulfoxide (DSSO) cross-linker. When disuccinimidyl suberate (DSS) cross-linker is in use, we are able to boost cross-link identification by 89% for the medium and 100% for the high complex sample compared to the analyses without FAIMS. Furthermore, we show that, for medium complex samples, FAIMS enables the collection of single-shot XL-MS data with a comparable depth to the corresponding sample fractionated by chromatography-based approaches. Altogether, we demonstrate FAIMS is highly beneficial for XL-MS studies by expanding the proteome coverage of cross-links while improving the efficiency and confidence of cross-link identification. |
| المشرفين على المادة: |
0 (Peptides) |
| تواريخ الأحداث: |
Date Created: 20200710 Date Completed: 20210222 Latest Revision: 20210222 |
| رمز التحديث: |
20240628 |
| DOI: |
10.1021/acs.analchem.0c01273 |
| PMID: |
32643919 |
| قاعدة البيانات: |
MEDLINE |
Find this issue from the American Chemical Society