We are planning to use Atomic Force Microscopy (AFM) to obtain information on the conformation of the membrane channels in the different condition, such as for instance in their native state, as well under the action of the different stimuli for the gating of the channel. To this purpose, we will exploit the high-resolution imaging capabilities in physiological conditions of AFM to study pore-containing natural and reconstituted phospholipid patches. We will develop suitable sample preparation protocols according to the specificity of the proteins involved. We will also tackle the challenging task of preparing the samples in a way that is suitable for the concurrent application of an external stimulus for the channel gating and the AFM investigation. In particular, we will consider special experimental setups allowing the application to the sample of controlled magnetic fields (using calibrated small magnets), IR fields (using infrared point sources), or ultrasound (by direct ultrasonic excitation of the sample-supporting substrate).
High-resolution Magic-Angle Spinning (HRMAS) NMR spectroscopy can be a powerful tool in facilitating to solve the chemical shift anisotropy problems and can also help in reducing the artefacts related to the presence of the superparamagnetic ferrihydrite-like mineral core of ferritin. Although we are aware that the presence of inhomogeneous size iron nanoparticles will prevent the acquisition of fine 2D resonance spectra, HRMAS DOSY experiments will be acquired to get relative information on the diffusion rate of ferritin in different media.
Our goal is to test in vivo the expression of the newly designed synthetic channels, whose activity should be controlled by diverse external stimuli (e.g. blue light, magnetic field). We exploit the zebrafish model system, taking advantage of its rapid embryonic and larval development and of its amenability to heterologous expression.
People in the lab involved in the project: Silvia MOLERI, post-doctoral researcher; Alessia BRIX, postgraduate degree level fellow
Our goal is the development and enhancement of in silico methods to understand and later on engineer the mechanics of ion channels and their signal-responsive domains. This is achieved by a combination of sequence analysis techniques and coarse-grained molecular models.
People in the lab involved in the project: Christine Gross, Daniel Bauer, Philipp Babel