Time-Resolved Tracking of Separately Internalized Neuropeptide Y Receptors by Two-Color Pulse-Chase.

Internalization and intracellular trafficking of G protein-coupled receptors (GPCR) plays an important role in the signal transduction. These processes are often highly dynamic and take place rapidly. In the past 10 years, it became obvious that internalized GPCRs are also capable of signaling via arrestin or heterotrimeric G proteins within the endosomal compartment.

Peptide-tags for site-specific protein labelling in vitro and in vivo.

Peptide-tag based labelling can be achieved by (i) enzymes (ii) recognition of metal ions or small molecules and (iii) peptide-peptide interactions and enables site-specific protein visualization to investigate protein localization and trafficking.

Rapid Covalent Fluorescence Labeling of Membrane Proteins on Live Cells via Coiled-Coil Templated Acyl Transfer.

Fluorescently labeled proteins enable the microscopic imaging of protein localization and function in live cells. In labeling reactions targeted against specific tag sequences, the size of the fluorophore-tag is of major concern. The tag should be small to prevent interference with protein function.

Peptide-templated acyl transfer: a chemical method for the labeling of membrane proteins on live cells.

The development of a method is described for the chemical labeling of proteins which occurs with high target specificity, proceeds within seconds to minutes, and offers a free choice of the reporter group. The method relies upon the use of peptide templates, which align a thioester and an N-terminal cysteinyl residue such that an acyl transfer reaction is facilitated at nanomolar concentrations. The protein of interest is N-terminally tagged with a 22 aa long Cys-E3 peptide (acceptor), which is capable of forming a coiled-coil with a reporter-armed K3 peptide (donor).

Experimental test of the spin mixing interface conductivity concept.

We perform a quantitative, comparative study of the spin pumping, spin Seebeck, and spin Hall magnetoresistance effects, all detected via the inverse spin Hall effect in a series of over 20 yttrium iron garnet/Pt samples. Our experimental results fully support present, exclusively spin current-based, theoretical models using a single set of plausible parameters for spin mixing conductance, spin Hall angle, and spin diffusion length. Our findings establish the purely spintronic nature of the aforementioned effects and provide a quantitative description, in particular, of the spin Seebeck effect.