Site-selective protein modification via disulfide rebridging for fast tetrazine/trans-cyclooctene bioconjugation

An inverse electron demand Diels–Alder reaction between tetrazine and trans-cyclooctene (TCO) holds great promise for protein modification and manipulation. Herein, we report the design and synthesis of a tetrazine-based disulfide rebridging reagent, which allows the site-selective installation of a tetrazine group into disulfide-containing peptides and proteins such as the hormone somatostatin (SST) and the antigen binding fragment (Fab) of human immunoglobulin G (IgG). The fast and efficient conjugation of the tetrazine modified proteins with three different TCO-containing substrates to form a set of bioconjugates in a site-selective manner was successfully demonstrated for the first time. Homogeneous, well-defined bioconjugates were obtained underlining the great potential of our method for fast bioconjugation in emerging protein therapeutics. The formed bioconjugates were stable against glutathione and in serum, and they maintained their secondary structure. With this work, we broaden the scope of tetrazine chemistry for site-selective protein modification to prepare well-defined SST and Fab conjugates with preserved structures and good stability under biologically relevant conditions

Electrically detected magnetic resonance of carbon dangling bonds at the Si-face 4H-SiC/SiO2_2 interface

SiC based metal-oxide-semiconductor field-effect transistors (MOSFETs) have gained a significant importance in power electronics applications. However, electrically active defects at the SiC/SiO$_2$ interface degrade the ideal behavior of the devices. The relevant microscopic defects can be identified by electron paramagnetic resonance (EPR) or electrically detected magnetic resonance (EDMR). This helps to decide which changes to the fabrication process will likely lead to further increases of device performance and reliability. EDMR measurements have shown very similar dominant hyperfine (HF) spectra in differently processed MOSFETs although some discrepancies were observed in the measured $g$-factors. Here, the HF spectra measured of different SiC MOSFETs are compared and it is argued that the same dominant defect is present in all devices. A comparison of the data with simulated spectra of the C dangling bond (P$_\textrm{bC}$) center and the silicon vacancy (V$_\textrm{Si}$) demonstrates that the P$_\textrm{bC}$ center is a more suitable candidate to explain the observed HF spectra.Comment: Accepted for publication in the Journal of Applied Physic

Subcellular mRNA localisation at a glance.

mRNA localisation coupled to translational regulation provides an important means of dictating when and where proteins function in a variety of model systems. This mechanism is particularly relevant in polarised or migrating cells. Although many of the models for how this is achieved were first proposed over 20 years ago, some of the molecular details are still poorly understood. Nevertheless, advanced imaging, biochemical and computational approaches have started to shed light on the cis-acting localisation signals and trans-acting factors that dictate the final destination of localised transcripts. In this Cell Science at a Glance article and accompanying poster, we provide an overview of mRNA localisation, from transcription to degradation, focusing on the microtubule-dependent active transport and anchoring mechanism, which we will use to explain the general paradigm. However, it is clear that there are diverse ways in which mRNAs become localised and target protein expression, and we highlight some of the similarities and differences between these mechanisms.This work was supported by a Wellcome Trust Senior Research Fellowship to I.D. supporting R.M.P. [grant number: 096144], a studentship from the Wellcome Trust to A.D. [grant number: 097304], the University of Cambridge, ISSF to T.T.W. [grant number 097814].This is the final version of the article. It first appeared from the Company of Biologists via http://dx.doi.org/10.1242/jcs.11427