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Multiplexed imaging mass spectrometry of the extracellular matrix using serial enzyme digests from formalin-fixed paraffin-embedded tissue sections.
Clift, Cassandra L; Drake, Richard R; Mehta, Anand; Angel, Peggi M.
Affiliation
  • Clift CL; Department of Cell and Molecular Pharmacology and Experimental Therapeutics, Medical University of South Carolina, 173 Ashley Avenue, Charleston, SC, 29425, USA.
  • Drake RR; Department of Cell and Molecular Pharmacology and Experimental Therapeutics, Medical University of South Carolina, 173 Ashley Avenue, Charleston, SC, 29425, USA.
  • Mehta A; Department of Cell and Molecular Pharmacology and Experimental Therapeutics, Medical University of South Carolina, 173 Ashley Avenue, Charleston, SC, 29425, USA.
  • Angel PM; Department of Cell and Molecular Pharmacology and Experimental Therapeutics, Medical University of South Carolina, 173 Ashley Avenue, Charleston, SC, 29425, USA. angelp@musc.edu.
Anal Bioanal Chem ; 413(10): 2709-2719, 2021 Apr.
Article in En | MEDLINE | ID: mdl-33206215
We report a multiplexed imaging mass spectrometry method which spatially localizes and selectively accesses the extracellular matrix on formalin-fixed paraffin-embedded tissue sections. The extracellular matrix (ECM) consists of (1) fibrous proteins, post-translationally modified (PTM) via N- and O-linked glycosylation, as well as hydroxylation on prolines and lysines, and (2) glycosaminoglycan-decorated proteoglycans. Accessing all these components poses a unique analytical challenge. Conventional peptide analysis via trypsin inefficiently captures ECM peptides due to their low abundance, intra- and intermolecular cross-linking, and PTMs. In previous studies, we have developed matrix-assisted laser desorption ionization imaging mass spectrometry (MALDI-IMS) techniques to capture collagen peptides via collagenase type III digestion, both alone and after N-glycan removal via PNGaseF digest. However, in fibrotic tissues, the buildup of ECM components other than collagen-type proteins, including elastin and glycosaminoglycans, limits efficacy of any single enzyme to access the complex ECM. Here, we have developed a novel serial enzyme strategy to define the extracellular matrix, including PTMs, from a single tissue section for MALDI-IMS applications. Graphical Abstract.
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Full text: 1 Collection: 01-internacional Database: MEDLINE Main subject: Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization / Extracellular Matrix Limits: Humans Language: En Journal: Anal Bioanal Chem Year: 2021 Document type: Article Affiliation country: Estados Unidos Country of publication: Alemania

Full text: 1 Collection: 01-internacional Database: MEDLINE Main subject: Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization / Extracellular Matrix Limits: Humans Language: En Journal: Anal Bioanal Chem Year: 2021 Document type: Article Affiliation country: Estados Unidos Country of publication: Alemania