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J Proteome Res ; 7(4): 1683-92, 2008 Apr.
Article in English | MEDLINE | ID: mdl-18338859

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.


Subject(s)
Cell Membrane Permeability/drug effects , Cell Membrane/drug effects , Proteins/analysis , Proteomics/methods , Saponins/pharmacology , Animals , Cell Line, Tumor , Cell Membrane/chemistry , Cell Membrane/ultrastructure , Cell Shape/drug effects , Cell Size/drug effects , Cholesterol/analysis , Chromatography, High Pressure Liquid/methods , Cytosol/metabolism , Electrophoresis, Gel, Two-Dimensional , Extracellular Space/chemistry , Hemolysis/drug effects , Humans , Hydro-Lyases/analysis , Micelles , Microscopy, Electron, Scanning , Microscopy, Phase-Contrast , Molecular Structure , Phospholipids/analysis , Saponins/chemistry , Sheep , Spectrometry, Mass, Secondary Ion , Tandem Mass Spectrometry
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