![]() Study membrane proteins in lipid nanoparticlesĮxtraction of membrane proteins from their native lipid environment with detergents often leads to the loss of structurally and functionally important lipids. Because the measurements are performed in solution, they have an advantage over other methods that require immobilization to a solid matrix. Studying membrane proteins in lipid nanodiscs and SMALPs keeps them in a membrane-like environment where interrogation of their structure and function is possible with methods like nanoDSF and MST. To characterize membrane proteins, researchers start by removing them from their native lipid environment - but this can fundamentally change their structure, and therefore their function. In combination with crystallography data, they revealed a key amino acid residue for nitrate binding and showed how phosphorylation of the transporter allows it to switch binding states. To understand this mechanism, researchers used MST to determine how nitrate binding to GFP-fused NRT1.1 in detergent was affected by phosphorylation and point mutations. In nature, it switches between low and high affinity nitrate binding states in response to falling nitrate levels via a process of phosphorylation. The NRT1.1 proton-coupled transporter is responsible for nitrogen assimilation and nitrate transport in plants. Identify key ligand binding sites in solution Additionally, they looked at how nanobodies stabilized the transporter for crystallization and structure determination with label-free nanoDSF. The data helped them model and improve drug design for the human transporter. Using DtpA - a bacterial homolog - this team measured the binding of peptides and drugs to DtpA with label-free MST. The human PepT1 transporter plays an important role in drug uptake and transport - which makes it a promising pharmacological target. Improve drug design to target membrane proteins The characterization of the interactions with ligands during drug discovery and development with label-free assays and in solution allows researchers to unravel molecular mechanisms and improve drug design. This knowledge also helps in designing drugs that target these processes. Study challenging interactions with membrane proteins to improve drug designįiguring out the role a membrane protein plays in a cellular process requires understanding how it interacts with its ligands. This team used nanoDSF as a tool for rapid screening of even very viscous LCP conditions. Just like with buffers and detergents, different types of LCP and additives are screened. The in meso crystallization method uses the lipid cubic phase (LCP) as a membrane-like environment to crystallize the proteins. Membrane proteins are difficult to crystallize because it’s challenging to solubilize them and maintain their native structure outside the membrane environment. Screen very viscous additives prior to crystallization The detergents that preserved the proteins’ stability were then used in purification and structural studies. This study applied label-free nanoDSF to rapidly screen 94 detergents by looking at the thermal stability of 9 membrane proteins. But finding the detergent that both solubilizes and preserves stability is difficult. Structural and functional characterization of membrane proteins starts with extracting the protein from its native membrane environment using detergents. Quickly find detergents that stabilize your membrane proteins Here are two examples that show how NanoTemper tools enable fast screening for detergents and viscous additives. To do this, researchers start by extracting enough protein from its native membrane, then screen different detergents and lipids to look for the optimal conditions that keep the protein folded and stable. Structure determination is a very important step in drug design for membrane proteins. ![]()
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