دورية أكاديمية
أعرض في EDS

Automated single-cell proteomics providing sufficient proteome depth to study complex biology beyond cell type classifications.

التفاصيل البيبلوغرافية
العنوان: Automated single-cell proteomics providing sufficient proteome depth to study complex biology beyond cell type classifications.
المؤلفون: Ctortecka C; Broad Institute of MIT and Harvard, Cambridge, MA, USA. cctortec@broadinstitute.org., Clark NM; Broad Institute of MIT and Harvard, Cambridge, MA, USA., Boyle BW; Broad Institute of MIT and Harvard, Cambridge, MA, USA., Seth A; Cellenion SASU, Lyon, France., Mani DR; Broad Institute of MIT and Harvard, Cambridge, MA, USA., Udeshi ND; Broad Institute of MIT and Harvard, Cambridge, MA, USA., Carr SA; Broad Institute of MIT and Harvard, Cambridge, MA, USA. scarr@broad.mit.edu.
المصدر: Nature communications [Nat Commun] 2024 Jul 08; Vol. 15 (1), pp. 5707. Date of Electronic Publication: 2024 Jul 08.
نوع المنشور: Journal Article
اللغة: English
بيانات الدورية: Publisher: Nature Pub. Group Country of Publication: England NLM ID: 101528555 Publication Model: Electronic Cited Medium: Internet ISSN: 2041-1723 (Electronic) Linking ISSN: 20411723 NLM ISO Abbreviation: Nat Commun Subsets: MEDLINE
أسماء مطبوعة: Original Publication: [London] : Nature Pub. Group
مواضيع طبية MeSH: Single-Cell Analysis*/methods , Proteomics*/methods , Proteome*/metabolism, Humans ; HEK293 Cells ; Ubiquitin-Protein Ligases/metabolism ; Mass Spectrometry/methods
مستخلص: The recent technological and computational advances in mass spectrometry-based single-cell proteomics have pushed the boundaries of sensitivity and throughput. However, reproducible quantification of thousands of proteins within a single cell remains challenging. To address some of those limitations, we present a dedicated sample preparation chip, the proteoCHIP EVO 96 that directly interfaces with the Evosep One. This, in combination with the Bruker timsTOF demonstrates double the identifications without manual sample handling and the newest generation timsTOF Ultra identifies up to 4000 with an average of 3500 protein groups per single HEK-293T without a carrier or match-between runs. Our workflow spans 4 orders of magnitude, identifies over 50 E3 ubiquitin-protein ligases, and profiles key regulatory proteins upon small molecule stimulation. This study demonstrates that the proteoCHIP EVO 96-based sample preparation with the timsTOF Ultra provides sufficient proteome depth to study complex biology beyond cell-type classifications.
(© 2024. The Author(s).)
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معلومات مُعتمدة: P01 CA206978 United States CA NCI NIH HHS; U01 CA271402 United States CA NCI NIH HHS; U24 CA270823 United States CA NCI NIH HHS; U24 CA271075 United States CA NCI NIH HHS
المشرفين على المادة: 0 (Proteome)
EC 2.3.2.27 (Ubiquitin-Protein Ligases)
تواريخ الأحداث: Date Created: 20240708 Date Completed: 20240708 Latest Revision: 20240722
رمز التحديث: 20240722
مُعرف محوري في PubMed: PMC11231172
DOI: 10.1038/s41467-024-49651-w
PMID: 38977691
قاعدة البيانات: MEDLINE
الوصف
تدمد:2041-1723
DOI:10.1038/s41467-024-49651-w