Pump-signalosome reconstituted in nano discs and small vesicles

Reconstitution of transmembrane proteins in nanodiscs.

We are trying to reconstitute the plasma membrane H⁺-pump in nano-discs. Nano discs are formed by a scaffolding protein that wrap around a lipid bilayer forming a "molecular sushi".

 

A receptor kinase regulating the activity of the plasma membrane H⁺-ATPase will also be expressed in yeast and reconstituted in nanodiscs and small vesicles. Thereby we can study the ligand-receptor interactions.

 

Finally the H⁺-pump together with the receptor kinase will be reconstituted into nanodiscs. The nanodiscs will be studied in solution by small angle x-ray scattering in the absence and the presence of receptor ligand in order to test whether perception of the signal causes structural changes in the signalosome complex (collaboration with Lise Arleth, Department of Basic Science).

 

 

A membrane scaffolding protein (MSP) folded around a lipid bilayer and a trans membrane protein (in our case the proton pump or the receptor kinase). Interacting proteins binding to the membrane protein is indicated as adaptor proteins. These can be regulatory proteins important for the function of the membrane protein (protein kinases or a ligand for the receptor).

 

This project is part of the UNIK programme Synthetic Biology and PUMPKIN - a Centre of Excellence funded by the Danish National Research Foundation.