TY - JOUR

T1 - Glyco-Steroidal Amphiphiles (GSAs) for Membrane Protein Structural Study

AU - Ehsan, Muhammad

AU - Wang, Haoqing

AU - Katsube, Satoshi

AU - Munk, Chastine F.

AU - Du, Yang

AU - Youn, Taeyeol

AU - Yoon, Soyoung

AU - Byrne, Bernadette

AU - Loland, Claus J.

AU - Guan, Lan

AU - Kobilka, Brian K.

AU - Chae, Pil Seok

N1 - Publisher Copyright: © 2022 Wiley-VCH GmbH

PY - 2022

Y1 - 2022

N2 - Integral membrane proteins pose considerable challenges to high resolution structural analysis. Maintaining membrane proteins in their native state during protein isolation is essential for structural study of these bio-macromolecules. Detergents are the most commonly used amphiphilic compounds for stabilizing membrane proteins in solution outside a lipid bilayer. We previously introduced a glyco-diosgenin (GDN) detergent that was shown to be highly effective at stabilizing a wide range of membrane proteins. This steroidal detergent has additionally gained attention due to its compatibility with membrane protein structure study via cryo-EM. However, synthetic inconvenience limits widespread use of GDN in membrane protein study. To improve its synthetic accessibility and to further enhance detergent efficacy for protein stabilization, we designed a new class of glyco-steroid-based detergents using three steroid units: cholestanol, cholesterol and diosgenin. These new detergents were efficiently prepared and showed marked efficacy for protein stabilization in evaluation with a few model membrane proteins including two G protein-coupled receptors. Some new agents were not only superior to a gold standard detergent, DDM (n-dodecyl-β-d-maltoside), but were also more effective than the original GDN at preserving protein integrity long term. These agents represent valuable alternatives to GDN, and are likely to facilitate structural determination of challenging membrane proteins.

AB - Integral membrane proteins pose considerable challenges to high resolution structural analysis. Maintaining membrane proteins in their native state during protein isolation is essential for structural study of these bio-macromolecules. Detergents are the most commonly used amphiphilic compounds for stabilizing membrane proteins in solution outside a lipid bilayer. We previously introduced a glyco-diosgenin (GDN) detergent that was shown to be highly effective at stabilizing a wide range of membrane proteins. This steroidal detergent has additionally gained attention due to its compatibility with membrane protein structure study via cryo-EM. However, synthetic inconvenience limits widespread use of GDN in membrane protein study. To improve its synthetic accessibility and to further enhance detergent efficacy for protein stabilization, we designed a new class of glyco-steroid-based detergents using three steroid units: cholestanol, cholesterol and diosgenin. These new detergents were efficiently prepared and showed marked efficacy for protein stabilization in evaluation with a few model membrane proteins including two G protein-coupled receptors. Some new agents were not only superior to a gold standard detergent, DDM (n-dodecyl-β-d-maltoside), but were also more effective than the original GDN at preserving protein integrity long term. These agents represent valuable alternatives to GDN, and are likely to facilitate structural determination of challenging membrane proteins.

KW - detergent design

KW - detergent-detergent interactions

KW - glyco-steroids

KW - glycolipids

KW - protein stabilization

U2 - 10.1002/cbic.202200027

DO - 10.1002/cbic.202200027

M3 - Journal article

C2 - 35129249

AN - SCOPUS:85124910064

VL - 23

JO - ChemBioChem

JF - ChemBioChem

SN - 1439-4227

IS - 7

M1 - e202200027

ER -