Overexpression and purification of MSP2N2 for nanodisc assembly and reconstitution of TRPA1

Sammanfattning: Membrane proteins are important building blocks for several biochemical processes. The difficulty with membrane proteins is that they tend to be lowly expressed and can be difficult to manage. Therefore, membrane proteins are studied to a lesser extent than soluble proteins. Since membrane proteins are intrinsically hydrophobic, their purification and structural determination critically rely on solubilization by detergents or other alternative procedures, such as nanodiscs. The purpose of this project is to overexpress and purify a membrane scaffold protein (MSP2N2) to allow assembly of nanodiscs, which in turn can help determine the structure of a membrane protein, namely a Transient Receptor Potential cation channel A1 (TRPA1) from Hylobius abietis. The initial results presented in this study shows the difficulty of producing MSP2N2 in terms of contaminants and degradation. These findings provide development opportunities for how to successfully isolate MSP2N2 for further use.