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Pores formation on cell membranes by hederacolchiside A1 leads to a rapid release of proteins for cytosolic subproteome analysis
Authors:Mazzucchelli Gabriel D  Cellier Nicolas A  Mshviladzade Vakhtang  Elias Riad  Shim Yong-Ho  Touboul David  Quinton Loïc  Brunelle Alain  Laprévote Olivier  De Pauw Edwin A  De Pauw-Gillet Marie-Claire A
Affiliation:Laboratory of Histology-Cytology (GIGA-CRCE), Institute of Chemistry (B6c), University of Liège, B-4000 Liège 1 (Sart-Tilman), Belgium.
Abstract:Hederacolchiside A1 was used to progressively permeabilize the membrane of human melanoma MEL-5 cells. Holes formation was followed by Scanning Electron Microscopy and interaction of the saponin with cholesterol and phospholipids by TOF-SIMS. 2D-LC-MS/MS and 2D-SDS-PAGE show that the release of soluble proteins into serum-free culture media increases with time. This can lead to a new rapid and efficient strategy to analyze the cytosolic subproteome and it opens the door to get information from the cytosolic compartment for clinical proteomic studies.
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