Hybrid Detergents Facilitate Scalable Charge Reduction and Stabilize Membrane Proteins While Retaining Lipid Interactions.

Urner LH, Shutin D, Agasid MT, Robinson CV

Membrane protein complexes regulate many biological functions, yet they are incredibly challenging to handle. Detergents are a key enabling technology for purification and analysis as they can provide membrane proteins with controlled solubility, stability, and heterogeneity. However, the design criteria for detergents that enable both the purification and analysis of membrane proteins with their associated lipids remain unclear. Here, we leverage the modular nature of nonionic hybrid detergents to chemically tune molecular parameters that are important for maintaining native oligomeric states of membrane protein complexes and copurifying lipids. Tuning the hydrophilic-lipophilic balance (HLB), and the relative size of a nonionic head in proximity to a tetraethylene glycol headgroup, led to altered charge reduction, retention of protein oligomers and changed the relative degree of copurifying lipids from purification to analysis in a predictable manner. Our findings represent a significant step forward in the investigation of membrane proteins and interactions with lipids.

Keywords:

Polyethylene Glycols

,

Lipids

,

Membrane Proteins

,

Detergents

,

Solubility

,

Hydrophobic and Hydrophilic Interactions