Modular detergents tailor the purification and structural analysis of membrane proteins including G-protein coupled receptors

Leonhard H Urner,Kin-Kuan Hoi,Kevin Pagel,Denis Shutin,Marc-Philip Schweder,Fernando Gonçalves Almeida,Idlir Liko,Svenja Ehrmann,Rainer Haag,Joseph Gault,Jani Reddy Bolla,Hsin-Yung Yen,Carol V Robinson

doi:10.1038/s41467-020-14424-8

Abstract

Detergents enable the purification of membrane proteins and are indispensable reagents in structural biology. Even though a large variety of detergents have been developed in the last century, the challenge remains to identify guidelines that allow fine-tuning of detergents for individual applications in membrane protein research. Addressing this challenge, here we introduce the family of oligoglycerol detergents (OGDs). Native mass spectrometry (MS) reveals that the modular OGD architecture offers the ability to control protein purification and to preserve interactions with native membrane lipids during purification. In addition to a broad range of bacterial membrane proteins, OGDs also enable the purification and analysis of a functional G-protein coupled receptor (GPCR). Moreover, given the modular design of these detergents, we anticipate fine-tuning of their properties for specific applications in structural biology. Seen from a broader perspective, this represents a significant advance for the investigation of membrane proteins and their interactions with lipids.

Highlights

Detergents enable the purification of membrane proteins and are indispensable reagents in structural biology
The ability to tune a particular detergent for isolating large protein quantities, and at the same time preserving protein interactions to native membrane lipids during isolation, is a major goal in membrane protein research[4,5,6]
We have introduced a library of oligoglycerol detergents (OGDs) to structural biology, which enables the isolation of a broad range of membrane proteins and facilitates their subsequent native mass spectrometry (MS) analysis

Summary

Introduction

Detergents enable the purification of membrane proteins and are indispensable reagents in structural biology. In the case of AqpZ, we found that the relative protein concentrations obtained following extraction and purification with 1 and 2 were about two times higher than with DDM (Fig. 2b). Protein quantities obtained from the [G1] OGD regioisomer mixture 1 were about three times higher than from DDM, leading to a higher yield of refolded OmpT under comparable conditions (Supplementary Fig. 4).

Results

Conclusion