Pores formation on cell membranes by hederacolchiside A1 leads to a rapid release of proteins for cytosolic subproteome analysis.
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.
Mazzucchelli
Gabriel D
GD
Laboratory of Histology-Cytology (GIGA-CRCE), Institute of Chemistry (B6c), University of Liège, B-4000 Liège 1 (Sart-Tilman), Belgium.
Cellier
Nicolas A
NA
Mshviladzade
Vakhtang
V
Elias
Riad
R
Shim
Yong-Ho
YH
Touboul
David
D
Quinton
Loïc
L
Brunelle
Alain
A
Laprévote
Olivier
O
De Pauw
Edwin A
EA
De Pauw-Gillet
Marie-Claire A
MC
D000818:Animals; D045744:Cell Line, Tumor; D002462:Cell Membrane / Q000737:chemistry / Q000187:drug effects* / Q000648:ultrastructure; D002463:Cell Membrane Permeability / Q000187:drug effects*; D048430:Cell Shape / Q000187:drug effects; D048429:Cell Size / Q000187:drug effects; D002784:Cholesterol / Q000032:analysis; D002851:Chromatography, High Pressure Liquid / Q000379:methods; D003600:Cytosol / Q000378:metabolism; D015180:Electrophoresis, Gel, Two-Dimensional; D005110:Extracellular Space / Q000737:chemistry; D006461:Hemolysis / Q000187:drug effects; D006801:Humans; D006836:Hydro-Lyases / Q000032:analysis; D008823:Micelles; D008855:Microscopy, Electron, Scanning; D008858:Microscopy, Phase-Contrast; D015394:Molecular Structure; D010743:Phospholipids / Q000032:analysis; D011506:Proteins / Q000032:analysis*; D040901:Proteomics / Q000379:methods*; D012503:Saponins / Q000737:chemistry / Q000494:pharmacology*; D012756:Sheep; D018629:Spectrometry, Mass, Secondary Ion; D053719:Tandem Mass Spectrometry
D008823:Micelles; D010743:Phospholipids; D011506:Proteins; D012503:Saponins; C467827:hederacolchisid A1; D002784:Cholesterol; D006836:Hydro-Lyases; C045581:lactate dehydratase