Abstract
Many cells respond to fluid shear stress. For example, endothelial cells produce vasodilator nitric oxide (NO) in response to blood flow. This response requires the presence of glypican-1, an extracellular GPI-anchoring protein; but the mechanism of activation is unknown. In environments like the circulatory system, there is some evidence that flow transport of extracellular lipid-anchored protein occurs. The problem is that it is unknown whether this transport contributes to flow sensing and signaling. Our lab developed a method to measure lipid-anchored protein mobility under flow in a simplified model system made up of supported bilayer patches formed from giant unilamellar vesicles (GUVs). However, physiologically interesting proteins obtained directly from cells via plasma membrane vesicles (PMVs) cannot be investigated using this method, since they end up oriented toward the solid support. Here, we demonstrate a procedure for creating finite supported membrane patches using microcontact printing with bovine serum albumin. This method results in patches with upper leaflets formed from the outer leaflets of the vesicles, making it possible to assay flow mobility of PMV-derived proteins. We show that we can measure protein flow mobility using this system. This will make it possible to compare mobility of the same protein in cells and in simplified supported membranes.
Talk to us
Join us for a 30 min session where you can share your feedback and ask us any queries you have
Schedule a callDisclaimer: All third-party content on this website/platform is and will remain the property of their respective owners and is provided on "as is" basis without any warranties, express or implied. Use of third-party content does not indicate any affiliation, sponsorship with or endorsement by them. Any references to third-party content is to identify the corresponding services and shall be considered fair use under The CopyrightLaw.
