SEC-SANS for investigation of three challenging and biologically relevant membrane proteins
We propose to utilize the SEC-SANS setup at D22 to investigate the three challenging and biologically relevant membrane proteins; growth hormone receptor (GHR), sodium/hydrogen exchanger1 (NHE1) and tissue factor (TF). All three proteins are expressed and purified in our lab, and solubilised in relevant car-rier systems. NHE1 will be investigated in match-out deuterated DDM, which will enable modeling of the protein structure without signal from the detergent micelle. GHR will be investigated in nanodiscs since the long intrinsically disordered intracellular domain of GHR might show a non-native interaction with its extracellular domain of when in a micelle. The nanodisc will also facilitate the investigation of the dimeri-zation and the interaction with growth hormone of the GHR. TF and the interaction with factor VIIa and factor Xa is highly dependent on the presence of a lipid bilayer and will hence also be studied in nano-discs. The SEC-SANS setup at D22 is a key factor in this approach, since the systems under investigation are highly delicate, and thus prone to form aggregates.
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ARLETH Lise; BLEMMER Sara; Nicolai Tidemand Johansen; KASSEM Noah; Andreas Haahr Larsen; LYCKSELL Marie; MARTEL Anne; Simone Orioli; PORCAR Lionel and F.G. Tidemand. (2019). SEC-SANS for investigation of three challenging and biologically relevant membrane proteins. Institut Laue-Langevin (ILL) doi:10.5291/ILL-DATA.8-03-959
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