The Electrofusion Mechanism in Erythrocyte Ghost Membranes
Abstract
Fusion of human erythrocyte ghost membranes was studied by using FITC-dextran and 1,1'-dihexadecyl-3,3,3',3'-tetramethylindo carbocyanine perchlorate (DiI) to detect, respectively, individual contents mixing events and membrane mixing events in populations of human erythrocytes. An electric field pulse was used as a fusogen and dielectrophoresis was used to reversibly induce membrane-membrane contact. We found during the first year of this project that when human erythrocyte ghosts in 20 mM or 60 mM sodium phosphate (pH 8.5) are treated with an appropriate electric field pulse to induce membrane fusion both fusion-associated contents mixing and non-fusion contents mixing are observed. In the present study separate assays conducted on the membrane preparation under identical conditions suggest that: i) the non-fusion contents mixing events are an artifact due to electroporation, ii) at 20 mM, fusion-associated contents mixing events (calculated by subtracting the fraction of non-fusion events) compare favorably with membrane mixing events over a wide range of fusion yields, iii) at 60 mM the non-fusion contents mixing events are suppressed, but interfering processes cause the fusion-associated contents mixing events to not compare favorably with membrane mixing events, and iv) electropores are not likely to be a fusion intermediate in the fusion mechanism. Keywords: Bioelectromagnetics.
Document Details
- Document Type
- Technical Report
- Publication Date
- Nov 30, 1988
- Accession Number
- ADA203041
Entities
People
- Arthur E. Sowers