Morth Group: Structural Facilities
A variety of techniques are used in order to identify, and characterise, both soluble and membrane bound proteins involved in metal/pH regulation.
- Structural modelling and bioinformatics approach are used to target new proteins and interaction partners of interest.
- A broad cloning strategy in both yeast and bacterial expression systems is employed to search for membrane/soluble proteins.
- Protocols for the purification and reconstitution of lipid vesicle for membrane proteins (Subramani, Perdreau-Dahl, and Morth, 2016)
- Structural information is obtained by X-ray crystallography, small angle X-ray scattering (SAXS) and small angle neutron scattering (SANS).
Several biophysical and biochemical techniques are also used:
- Fluorescent binding assays, including drug screening (Gerner, Munack, et al, 2016a; Gerner, Munack, et al, 2016b)
- Differential scanning fluorimetry (DSF), for melting point, folding and stability measures (Leo, Oberhettinger, et al, 2016)
- Anisotropic fluorescence measurement, stop flow
- Spectroscopic activity assays and enzyme kinetics (Bjerregaard-Andersen, Sommer, et al, 2014; Sommer, Bjerregaard-Andersen, et al, 2015; Subramani, Perdreau-Dahl, and Morth, 2016).
- Bjerregaard-Andersen, K., Sommer, T., Jensen, J.K., Jochimsen, B., Etzerodt, M., and Morth, J.P. (2014). A proton wire and water channel revealed in the crystal structure of isatin hydrolase. J. Biol. Chem.
- Gerner, L., Munack, S., Temmerman, K., Lawrence-Dorner, A.M., Besir, H., Wilmanns, M., Jensen, J.K., Thiede, B., Mills, I.G., and Morth, J.P. (2016a). Data for the co-expression and purification of human recombinant CaMKK2 in complex with calmodulin in Escherichia coli. Data Brief 733-740.
- Gerner, L., Munack, S., Temmerman, K., Lawrence-Dorner, A.M., Besir, H., Wilmanns, M., Jensen, J.K., Thiede, B., Mills, I.G., and Morth, J.P. (2016b). Using the fluorescent properties of STO-609 as a tool to assist structure-function analyses of recombinant CaMKK2. Biochem. Biophys. Res. Commun. 2, 102-107.
- Leo, J.C., Oberhettinger, P., Yoshimoto, S., Udatha, D.B., Morth, J.P., Schutz, M., Hori, K., and Linke, D. (2016). Secretion of the Intimin Passenger Domain is driven by Protein Folding. J. Biol. Chem.
- Sommer, T., Bjerregaard-Andersen, K., Simensen, S.M., Jensen, J.K., Jochimsen, B., Riss, P.J., Etzerodt, M., and Morth, J.P. (2015). Enzymatic Detection and Quantification Assay of Isatin, a Putative Stress Biomarker in Blood. ACS Chem. Neurosci.
- Subramani, S., Perdreau-Dahl, H., and Morth, J.P. (2016). The magnesium transporter A is activated by cardiolipin and is highly sensitive to free magnesium in vitro. Elife 10.7554/eLife.11407.