Emerging Concepts in Ion Channel Biophysics

Emerging Concepts in Ion Channel Biophysics

Wednesday Speaker Abstracts

Optical Approaches in Studies of Excitability

Francisco Bezanilla

University of Chicago, USA.

No Abstract

Minimal Tags for Live-Cell Protein Labelling and Superresolution Microscopy

Ivana Nikic-Spiegel

Werner Reichardt Centre for Integrative Neuroscience (CIN), Tuebingen, Germany.

Genetic code expansion (GCE) is emerging as an important technology for

in vitro

and

vivo

protein manipulation and labelling. In combination with click-chemistry it allows site-

specific labelling of proteins with small organic dyes. This is achieved by co-translational

incorporation of unnatural amino acids (UAAs) in target proteins by using tRNA/amino-acyl

tRNA synthetase pairs orthogonal to the host translational machinery. In a subsequent step,

unique functional groups of UAAs are labelled with functionalized dyes in ultrafast and

biocompatible click-chemistry reactions. The fact that any dye can be directly attached to the

target protein in a minimally invasive way is of particular importance for single molecule science

and superresolution microscopy (SRM). We previously used this technology for dual-colour live-

cell labelling and SRM of distinct populations of membrane proteins in mammalian cells. In our

current work we use it to visualize axonal injury in neuroinflammatory diseases, such as multiple

sclerosis, at a nanoscale level.