| Literature DB >> 33232116 |
Samantha J Knott1, Kyle A Brown1, Harini Josyer2, Austin Carr1, David Inman2, Song Jin1, Andreas Friedl3, Suzanne M Ponik2, Ying Ge1,2,4.
Abstract
The extracellular matrix (ECM) provides an architectural meshwork that surrounds and supports cells. The dysregulation of heavily post-translationally modified ECM proteins directly contributes to various diseases. Mass spectrometry (MS)-based proteomics is an ideal tool to identify ECM proteins and characterize their post-translational modifications, but ECM proteomics remains challenging owing to the extremely low solubility of the ECM. Herein, enabled by effective solubilization of ECM proteins using our recently developed photocleavable surfactant, Azo, we have developed a streamlined ECM proteomic strategy that allows fast tissue decellularization, efficient extraction and enrichment of ECM proteins, and rapid digestion prior to reversed-phase liquid chromatography (RPLC)-MS analysis. A total of 173 and 225 unique ECM proteins from mouse mammary tumors have been identified using 1D and 2D RPLC-MS/MS, respectively. Moreover, 87 (from 1DLC-MS/MS) and 229 (from 2DLC-MS/MS) post-translational modifications of ECM proteins, including glycosylation, phosphorylation, and hydroxylation, were identified and localized. This Azo-enabled ECM proteomics strategy will streamline the analysis of ECM proteins and promote the study of ECM biology.Entities:
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Year: 2020 PMID： 33232116 PMCID： PMC7961849 DOI： 10.1021/acs.analchem.0c03104
Source DB: PubMed Journal: Anal Chem ISSN： 0003-2700 Impact factor: 6.986