Physics (Internal)

Challenges associated with de novo synthesis of functional receptors supported in nanolipoproteins

This event is part of the Collider Search Candidate Seminars.

Abstract:
We have developed several cell-free methods for producing soluble membrane proteins supported in nanolipoprotein particles (NLPs) for the structural study of membrane proteins. NLPs have shown great promise as a biotechnology platform for solubilizing and characterizing membrane proteins and may make many more membrane proteins accessible for biophysical and biochemical study. By the simple addition of a few constituents to cell-free extracts, this approach provides a rapid process for the production of functional soluble membrane proteins that eliminates the need for cell growth, cell lysis, and subsequent purification, refolding etc. We have demonstrated this process on multiple membrane proteins important for rhodopsin-related proton pumps, drug transporters, host-pathogen interactions, the herceptin receptors and G-coupled protein receptors. Proteins have ranged in size from 10 – 200 kDa, with 2 or more transmembrane domains and have been shown to be biochemically functional. Our approach represents a unique solution to the inherent challenge of generating soluble and functional polytopic membrane proteins, facilitating the structural and functional characterization of these critical, yet poorly understood molecules.