Nanolipoprotein particles and related methods and systems for protein capture, solubilization, and/or purification
Provided herein are methods and systems for assembling, solubilizing and/or purifying a membrane associated protein in a nanolipoprotein particle, which comprise a temperature transition cycle performed in presence of a detergent, wherein during the temperature transition cycle the nanolipoprotein components are brought to a temperature above and below the gel to liquid crystalling transition temperature of the membrane forming lipid of the nanolipoprotein particle.
1. A method for assembling a membrane associated protein, a scaffold protein, and a membrane forming lipid into a nanolipoprotein particle, the membrane forming lipid having a membrane forming lipid gel-crystalline transition temperature, the method comprising
contacting the membrane associated protein with the scaffold protein and the membrane forming lipid to provide an admixture; and
subjecting the admixture to multiple temperature transition cycles in presence of a detergent, for a time and under condition to allow assembly of the nanolipoprotein particle, the temperature transition cycle comprising:
a temperature increase step wherein the admixture is brought to a temperature above said membrane forming lipid gel crystalline transition temperature and
a temperature decrease step wherein the admixture is brought to a temperature below said membrane forming lipid gel crystalline transition temperature.
2. The method of claim 1 , wherein the membrane forming lipid is contacted with the membrane associated protein and the scaffold protein at a temperature above the membrane forming lipid gel-crystalline transition temperature.
3. The method of claim 1 , wherein the temperature increase step and/or the temperature decrease step are performed for a time and under conditions such that at least a fraction of the membrane forming lipid molecules in the admixture changes state.
4. The method of claim 1 , wherein the membrane associated protein contacted with the membrane forming lipid and with the scaffold protein is comprised in a cell membrane.
5. The method of claim 1 , further comprising incubating the admixture subjected to the temperature transition cycle at the membrane forming lipid gel-crystalline transition temperature for a predetermined amount of time that is functional to a desired amount of assembled nanolipoprotein particles.
6. The method of claim 1 , further comprising removing the detergent from the admixture subjected to the temperature transition cycle.