Nanolipoprotein particles and related methods and systems for protein capture, solubilization, and/or purification

摘要

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.

主权项

1. A method for solubilizing a membrane associated protein from a cell membrane, the method comprising
contacting the cell membrane with a scaffold protein and a membrane forming lipid to provide an admixture, the membrane forming lipid having a membrane forming lipid gel-crystalline transition temperature; and subjecting the admixture to at least three temperature transition cycles in presence of a detergent, for a time and under condition to allow assembly of a nanolipoprotein particle comprising a solubilized target protein, 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.