High-Throughput Extracellular Matrix Proteomics of Human Lungs Enabled by Photocleavable Surfactant and diaPASEF

J Proteome Res. 2024 Aug 2;23(8):2908-2918. doi: 10.1021/acs.jproteome.3c00532. Epub 2024 Feb 5.

Abstract

The extracellular matrix (ECM) is a complex assembly of proteins that provide interstitial scaffolding and elastic recoil for human lungs. The pulmonary extracellular matrix is increasingly recognized as an independent bioactive entity, by creating biochemical and mechanical signals that influence disease pathogenesis, making it an attractive therapeutic target. However, the pulmonary ECM proteome ("matrisome") remains challenging to analyze by mass spectrometry due to its inherent biophysical properties and relatively low abundance. Here, we introduce a strategy designed for rapid and efficient characterization of the human pulmonary ECM using the photocleavable surfactant Azo. We coupled this approach with trapped ion mobility MS with diaPASEF to maximize the depth of matrisome coverage. Using this strategy, we identify nearly 400 unique matrisome proteins with excellent reproducibility that are known to be important in lung biology, including key core matrisome proteins.

Keywords: Azo; Data Independent Acquisition; Diapasef; Extracellular Matrix; Matrisome.

MeSH terms

  • Azo Compounds / chemistry
  • Extracellular Matrix Proteins / metabolism
  • Extracellular Matrix* / metabolism
  • Humans
  • Lung* / metabolism
  • Mass Spectrometry / methods
  • Proteome / analysis
  • Proteomics* / methods
  • Surface-Active Agents / chemistry

Substances

  • Azo Compounds
  • Extracellular Matrix Proteins
  • Surface-Active Agents
  • Proteome