New penta-saccharide-bearing tripod amphiphiles for membrane protein structure studies

Abstract

Integral membrane proteins either alone or as complexes carry out a range of key cellular functions. Detergents are indispensable tools in the isolation of membrane proteins from biological membranes for downstream studies. Although a large number of techniques and tools, including a wide variety of detergents, are available, purification and structural characterization of many membrane proteins remain challenging. In the current study, a new class of tripod amphiphiles bearing two different penta-saccharide head groups, designated TPSs, were developed and evaluated for their ability to extract and stabilize a range of diverse membrane proteins. Variations in the structures of the detergent head and tail groups allowed us to prepare three sets of the novel agents with distinctive structures. Some TPSs (TPS-A8 and TPS-E7) were efficient at extracting two proteins in a functional state while others (TPS-E8 and TPS-E10L) conferred marked stability to all membrane proteins (and membrane protein complexes) tested here compared to a conventional detergent. Use of TPS-E10L led to clear visualization of a receptor-Gs complex using electron microscopy, indicating profound potential in membrane protein research.

1,3,5-Triazine-Cored Maltoside Amphiphiles for Membrane Protein Extraction and Stabilization

“…This study varied both the alkyl chain linkage (ether/thioether) and linker identity (serinol/DEA), allowing us to investigate the effects of these structural differences on membrane protein stability in detail. In previous studies, an ether linkage has been compared with an alkyl linkage, ,, but it is unusual to compare the thioether linkage with another (ether or alkyl) in the context of novel detergent structure. Due to the high hydrophobicity of the sulfur atom (vs oxygen) and relatively long C–S bond (vs C–O), the thioether linkage-bearing detergents gave lower CMCs and larger micelle sizes than the ether counterparts, respectively (Table ).…”

Solution Properties of Alkyl β‐D‐Maltosides

“…However, there are not enough data and messages available to support effectively the application in the field of life and material sciences although there are some available features/parameters including critical micelle concentration (CMC) (Frotscher et al, 2017;Matsuoka et al, 2017;Santonicola et al, 2008), sugar headgroup structure (Ehsan et al, 2016), field-induced phase transition (Hashim et al, 2017), paracellular permeation (Gradauer et al, 2017), and cell metabolism (Smułek et al, 2017). The corresponding fairly accurate and feasible screening should rely heavily on the empirical evidence, exhaustive trial-and-error process and rational thinking, and even intuitive approach.…”

Steroid‐Based Amphiphiles for Membrane Protein Study: The Importance of Alkyl Spacers for Protein Stability

“…Of the SPSs and SPS‐Ls, SPS‐2L and SPS‐3L were the best, because these were capable of retaining quantitative amounts of soluble MelB St even at 75 °C. It is noteworthy that most new detergents developed to date have failed to yield noticeable amounts of soluble MelB St even at 65 °C under similar conditions . Therefore, these results indicate that the SPSs (SPS‐1/2/3) and SPS‐Ls (SPS‐1L/2L/3L) are remarkably effective at maintaining the transporter in a soluble state.…”