PMID: 11334966May 4, 2001Paper

Cytolytic peptides induce biphasic permeability changes in mammalian cell membranes

Journal of Immunological Methods
M SuS J Mentzer

Abstract

The cytolytic peptides melittin and gramicidin S are naturally occurring agents that provide a comparative model for studies of complement, immunotoxin and cell-mediated membrane permeability. Most attempts to characterize cytolytic peptides have used model membrane systems including phospholipid vesicles or erythrocytes. Membrane vesicles permit the use of self-quenching concentrations of fluorescent permeability markers, while erythrocytes release measurable hemoglobin. Attempts at measuring early membrane permeability changes in nucleated mammalian cells have been limited. To measure the kinetics of mammalian cell membrane permeability changes induced by cytolytic peptides, we developed a 96-well fluorescence cytolysis assay using the cytoplasmic fluorescent dye calcein as the membrane permeability marker. To facilitate rapid assessment of membrane permeability, trypan blue was added to the assay solution to quench (a) released fluorescence and (b) retained intracellular fluorescence. Trypan blue also provided a complementary visual assessment of cell viability. Using this assay, a detailed kinetic analysis demonstrated permeability of the cell membranes within seconds of exposure to the cytolytic peptides. The rapid permeab...Continue Reading

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Citations

Jun 30, 2005·BioDrugs : Clinical Immunotherapeutics, Biopharmaceuticals and Gene Therapy·Josias H HammanAwie F Kotzé
Feb 2, 2006·In Vitro Cellular & Developmental Biology. Animal·Juan Pablo PrattSteven J Mentzer
Jan 3, 2007·The Journal of Thoracic and Cardiovascular Surgery·Dino J RavnicSteven J Mentzer
Apr 20, 2011·Journal of Cellular Physiology·Barry C GibneySteven J Mentzer
Dec 2, 2006·Bioscience Reports·H Raghuraman, Amitabha Chattopadhyay
May 16, 2012·Molecular Membrane Biology·John M Sanderson
Oct 14, 2014·Frontiers in Oncology·Robert D BennettSteven J Mentzer
Apr 28, 2006·FEMS Microbiology Letters·Judy HigginsTimothy J Foster
Aug 13, 2005·Microvascular Research·Dino J RavnicMoritz A Konerding
Feb 13, 2018·The Journal of Clinical Investigation·Jaime L HookJahar Bhattacharya
Jul 4, 2019·Biomacromolecules·Trisha L BaileyMatthew I Gibson

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