Int J Biol Sci 2010; 6(7):649-654. doi:10.7150/ijbs.6.649 This issue

Short Research Communication

Mechanical Forces Used for Cell Fractionation Can Create Hybrid Membrane Vesicles

Izhar Salomon, Hans Janssen, Jacques Neefjes

The Netherlands Cancer Institute, Division of Cell Biology. Amsterdam, the Netherlands.

This is an open access article distributed under the terms of the Creative Commons Attribution (CC BY-NC) License. See http://ivyspring.com/terms for full terms and conditions.
Citation:
Salomon I, Janssen H, Neefjes J. Mechanical Forces Used for Cell Fractionation Can Create Hybrid Membrane Vesicles. Int J Biol Sci 2010; 6(7):649-654. doi:10.7150/ijbs.6.649. Available from https://www.ijbs.com/v06p0649.htm

File import instruction

Abstract

The ability to understand the inner works of the cell requires methods for separation of intracellular membrane-enclosed compartments. Disruption of the plasma membrane (PM) by mechanical forces to investigate the content of the cell is common practice. Whether vesicles or membranes of different sources can fuse as a result is unclear. If such contamination occurs, conclusions based on these techniques should consider these. Utilizing an endoplasmic reticulum (ER) membrane marker and a PM marker, we were able to detect the source of membranes following the breakup of cells using flow cytometry and immuno Electron Microscopy (immuno EM). Fractionation processes produced a small fraction of new membrane entities from two distinctively different origins generated during the initial disruption steps in a temperature independent manner, stressing that defining organelles or intrinsic fusion events based on such procedures and markers are valid when exceeding the small number of vesciles fused during the fractionation process.

Keywords: membrane fusion, fractionation, trafficking, signalling, proteomics.