Rationally Engineered Tandem Facial Amphiphiles for Improved Membrane Protein Stabilization Efficacy

Abstract

A new family of tandem facial glucosides/maltosides (TFGs/TFMs) for membrane protein manipulation was prepared. The best detergent varied depending on the hydrophobic thickness of the target protein, but ether-based TFMs (TFM-C0E, TFM-C3E, and TFM-C5E) were notable for their ability to confer higher membrane protein stability than the previously developed amide-based TFA-1 (P. S. Chae, K. Gotfryd, J. Pacyna, L. J. W. Miercke, S. G. F. Rasmussen, R. A. Robbins, R. R. Rana, C. J. Loland, B. Kobilka, R. Stroud, B. Byrne, U. Gether, S. H. Gellman, J. Am. Chem. Soc. 2010, 132, 16750-16752). Thus, this study not only introduces novel agents with the potential to be used in membrane protein research but also highlights the importance of both the hydrophobic length and linker functionality of the detergent in stabilizing membrane proteins.

Tris(hydroxymethyl)aminomethane Linker-Bearing Triazine-Based Triglucosides for Solubilization and Stabilization of Membrane Proteins

“…This alkyl chain length variation is necessary for the following two reasons. First, the detergent alkyl chain needs to be compatible with the hydrophobic dimensions of membrane proteins for protein stability. , Membrane proteins have a range of hydrophobic width (∼30 Å), and thus a specific alkyl chain length may be required depending on the exact hydrophobic width of a given membrane protein . Second, detergent alkyl chain length varies the hydrophilic–lipophilic balance (HLB) value, important for membrane protein stability .…”

“…21,22 Membrane proteins have a range of hydrophobic width (∼30 Å), and thus a specific alkyl chain length may be required depending on the exact hydrophobic width of a given membrane protein. 42 Second, detergent alkyl chain length varies the hydrophilic−lipophilic balance (HLB) value, important for membrane protein stability. 43 The HLB values of the TTGs calculated from Griffin's method are given in Table S1.…”

Methods for the solubilisation of membrane proteins: the micelle-aneous world of membrane protein solubilisation

“…However, recently there has been a drive towards developing completely novel detergents that confer, in particular, elevated protein stability, compared with classical detergents. Examples of these include; a new class of steroid-based pentasaccharides [ 6 ], fluorinated glucose and maltose-based detergents [ 7 , 8 ], innovative facial amphiphiles (FAs) [ 9 , 10 ], 1,3,5-triazine-cored detergents [ 11 ], disulfide containing amphiphiles [ 12 ] and modified neopentyl glycol based detergents that have undergone subsequent modification to further improve their characteristics [ 13 ] ( Figure 2 ).…”

Two‐Dimensional Detergent Expansion Strategy for Membrane Protein Studies

“…Similarly, amphipols are seemingly derived from the polymerization of detergent monomers. Facial amphiphiles (FAs) [27,42,49,50] result from the anchoring of hydrophilic heads on the cholane scaffold, and the dimerization of FAs yields tandem facial amphiphiles, which can be considered as a double preassembly. Peptide-based detergents, particularly β-sheet peptide detergents, [29] can be considered to derive from a wellstructured hydrogen bond-mediated assembly process involving the amphipathic β-strand, which itself is formed by a linear array of small detergents (Chart S1 in Supporting Information).…”

“…This approach results in a size‐limited pool of commercially available detergents that are far from sufficient for the discovery of optimal detergent for high‐fidelity sample preparation. During the past decades, new detergents featuring enlarged molecular sizes with multiple heads and multiple tails have emerged with continuous introduction of creative scaffolds (Chart S1 in Supporting Information) [26–48] . We designate these nontraditional detergents as pre‐assembled detergents (PADs) on the basis of topology.…”