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.
Publication types
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Research Support, Non-U.S. Gov't
MeSH terms
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Animals
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Cell Line, Tumor
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Cell Membrane / chemistry
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Cell Membrane / drug effects*
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Cell Membrane / ultrastructure
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Cell Membrane Permeability / drug effects*
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Cell Shape / drug effects
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Cell Size / drug effects
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Cholesterol / analysis
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Chromatography, High Pressure Liquid / methods
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Cytosol / metabolism
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Electrophoresis, Gel, Two-Dimensional
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Extracellular Space / chemistry
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Hemolysis / drug effects
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Humans
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Hydro-Lyases / analysis
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Micelles
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Microscopy, Electron, Scanning
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Microscopy, Phase-Contrast
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Molecular Structure
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Phospholipids / analysis
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Proteins / analysis*
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Proteomics / methods*
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Saponins / chemistry
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Saponins / pharmacology*
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Sheep
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Spectrometry, Mass, Secondary Ion
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Tandem Mass Spectrometry
Substances
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Micelles
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Phospholipids
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Proteins
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Saponins
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hederacolchisid A1
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Cholesterol
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Hydro-Lyases
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lactate dehydratase