title: Analytical characterization of plasma membrane-derived vesicles produced via osmotic and chemical vesiculation.
abstract: Plasma membrane-derived vesicles are being used in biophysical and biochemical research as a simple, yet native-like model of the cellular membrane. Here we report on the characterization of vesicles produced via two different vesiculation methods from CHO and A431 cell lines. The first method is a recently developed method which utilizes chloride salts to induce osmotic vesiculation. The second is a well established chemical vesiculation method which uses DTT and formaldehyde. We show that both vesiculation methods produce vesicles which contain the lipid species previously reported in the plasma membrane of these cell lines. The two methods lead to small but statistically significant differences in two lipid species only; phosphatidylcholine (PC) and plasmalogen phosphatidylethanolamine (PEp). However, highly significant differences were observed in the degree of incorporation of a membrane receptor and in the degree of retention of soluble cytosolic proteins within the vesicles.
authors.affiliation: Department of Materials Sciences and Engineering, Johns Hopkins University, Baltimore, MD 21218, USA.
authors.forename: Sarvenaz
authors.initials: S
authors.lastname: Sarabipour
authors.affiliation: Department of Pathology and Cell Biology, Columbia University Medical Center, Columbia University, New York, NY 10032, USA.
authors.forename: Robin B
authors.initials: RB
authors.lastname: Chan
authors.affiliation: Department of Pathology and Cell Biology, Columbia University Medical Center, Columbia University, New York, NY 10032, USA.
authors.forename: Bowen
authors.initials: B
authors.lastname: Zhou
authors.affiliation: Department of Pathology and Cell Biology, Columbia University Medical Center, Columbia University, New York, NY 10032, USA.
authors.forename: Gilbert
authors.initials: G
authors.lastname: Di Paolo
authors.affiliation: Department of Materials Sciences and Engineering, Johns Hopkins University, Baltimore, MD 21218, USA. Electronic address: kh@jhu.edu.
authors.forename: Kalina
authors.initials: K
authors.lastname: Hristova
mesh_terms: D000818:Animals; D015153:Blotting, Western; D016466:CHO Cells; D045744:Cell Line, Tumor; D002462:Cell Membrane / Q000737:chemistry / Q000378:metabolism*; D002784:Cholesterol / Q000378:metabolism; D002853:Chromatography, Liquid; D006224:Cricetinae; D003412:Cricetulus; D004229:Dithiothreitol / Q000494:pharmacology; D066246:ErbB Receptors / Q000378:metabolism; D005557:Formaldehyde / Q000494:pharmacology; D006801:Humans; D013058:Mass Spectrometry; D008563:Membrane Lipids / Q000378:metabolism*; D008565:Membrane Proteins / Q000378:metabolism*; D018613:Microscopy, Confocal; D009997:Osmotic Pressure; D010713:Phosphatidylcholines / Q000378:metabolism; D010714:Phosphatidylethanolamines / Q000378:metabolism; D010955:Plasmalogens / Q000378:metabolism; D022161:Transport Vesicles / Q000737:chemistry / Q000187:drug effects / Q000378:metabolism*
chemical_list: D008563:Membrane Lipids; D008565:Membrane Proteins; D010713:Phosphatidylcholines; D010714:Phosphatidylethanolamines; D010955:Plasmalogens; D005557:Formaldehyde; C483858:phosphatidylethanolamine; D002784:Cholesterol; D066246:ErbB Receptors; D004229:Dithiothreitol
keywords: Lipid composition; Membrane; Vesicle