12/19/2023 0 Comments Molecular scaffold protein![]() Supplied as a lyophilized histidine-tagged protein with a TEV protease cleavage site stabilized with Tris-HCl, EDTA, and NaCl. 7,575,763 Membrane scaffold proteins and tethered membrane proteins Mapping of genetic, proteomic and biochemical data onto known protein-protein and protein-RNA interaction maps has suggested that molecules with higher connectivity to other molecules in the condensate, and thus located more centrally in the condensate interaction network, are likely to behave more scaffold-like, while those with lower.7,622,437 Tissue factor compositions and methods.7,662,410 Membrane scaffold proteins and embedded membrane proteins.Nanodisc technology, and many of its uses, are covered by the following patents held by the University of Illinois. The thickness of a nanodisc is dependent on the type of phospholipid incorporated (typically 4.6−5.6 nm). Biophysical investigations of membrane proteins are noto- riously difficult. ![]() The length of the MSP controls the size of the nanodisc structure. Molecular & Cellular Proteomics 10: 10.1074/ mcp.M111.010876, 111, 2011. A soluble nanodisc assembles as the phospholipid forms a bilayer, which is encircled by two amphipathic MSP molecules covering the hydrophobic alkyl chains of the bilayer. Over the past decade, examples of scaffolding complexes long thought to be stable have instead been found to be surprisingly dynamic. Numerous scaffolding proteins are found in nature, many having multiple proteinprotein interaction modules. The nanodisc is a non-covalent structure of phospholipid bilayer and membrane scaffold protein (MSP), a genetically engineered protein, which mimics the function of Apolipoprotein A-1 (ApoA-1). Studies have revealed the central role of multivalent interactions in driving condensate formation 3,4.Many instantiations of multivalency have been described in condensates, involving folded protein domains, intrinsically disordered regions (IDR) 3,5, nucleic acids 3, and chromatin 6. The function of scaffolding proteins is to bring together two or more proteins in a relatively stable configuration, hence their name. The nanodisc concept is derived from high density lipoprotein (HDL) particles and their primary protein component, apolipoprotein. The nanodisc system has been employed to incorporate a wide variety of proteins including GPCRs, P450s, bacteriorhodopsin, coagulation factors, cholera toxin, TAR receptor and aromatase. The critical component of nanodiscs is the encircling amphipathic helical protein belt (membrane scaffold protein). Nanodisc soluble lipid bilayer systems have proven to be a widely applicable means for rendering membrane proteins soluble in aqueous solutions in a native-like bilayer environment where they remain monodisperse and active. 26,600 M -1cm -1 at 280 nm (for the His-tag-cleaved form dissolved in 20 mM Tris pH 7.4, 0.1M NaCl, 0.5mM EDTA and 0.01%NaN 3)(lit.)Ģ9,400 M -1cm -1 at 280 nm (for the uncleaved His-tagged form dissolved in 20 mM Tris pH 7.4, 0.1M NaCl, 0.5mM EDTA and 0.01%NaN 3)(lit.)įor guidelines on the use of this and other MSP′s to prepare Nanodiscs, please visit our Protocols for Membrane Scaffold Proteins and Nanodisc Formation page. ![]()
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