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Photocleavable Surfactant-Enabled Extracellular Matrix Proteomics. | LitMetric

Photocleavable Surfactant-Enabled Extracellular Matrix Proteomics.

Anal Chem

Department of Chemistry, University of Wisconsin-Madison, 1101 University Avenue, Madison, Wisconsin 53706, United States.

Published: December 2020

AI Article Synopsis

  • - The extracellular matrix (ECM) supports cells but its altered proteins can lead to diseases, making the study of ECM crucial for understanding health issues.
  • - Traditional methods to analyze ECM proteins have been challenging due to their low solubility; however, a new photocleavable surfactant called Azo has improved this process.
  • - Using this new strategy, researchers identified 173 and 225 unique ECM proteins from mouse mammary tumors, alongside various post-translational modifications, enhancing the study of ECM biology.

Article 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.

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Source
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7961849PMC
http://dx.doi.org/10.1021/acs.analchem.0c03104DOI Listing

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