EquipSent - Enabling Sustainable Education, Everywhere.

EquipSent is a volunteer-based non-profit organization aiming at creating conditions for sustainable teaching, study, and academic research worldwide. Used, functional equipment is collected by its members, who are responsible for matching the donations with the receivers in need. After starting in 2017, nine big transfers were accomplished that significantly impacted the quality of local scientific and educational life.

Dissecting the Molecular Origin of -Tensor Heterogeneity and Strain in Nitroxide Radicals in Water: Electron Paramagnetic Resonance Experiment versus Theory.

Nitroxides are common EPR sensors of microenvironmental properties such as polarity, numbers of H-bonds, pH, and so forth. Their solvation in an aqueous environment is facilitated by their high propensity to form H-bonds with the surrounding water molecules. Their - and -tensor elements are key parameters to extracting the properties of their microenvironment.

Spectroscopically Orthogonal Spin Labels in Structural Biology at Physiological Temperatures.

Electron paramagnetic resonance spectroscopy (EPR) is mostly used in structural biology in conjunction with pulsed dipolar spectroscopy (PDS) methods to monitor interspin distances in biomacromolecules at cryogenic temperatures both in vitro and in cells. In this context, spectroscopically orthogonal spin labels were shown to increase the information content that can be gained per sample. Here, we exploit the characteristic properties of gadolinium and nitroxide spin labels at physiological temperatures to study side chain dynamics via continuous wave (cw) EPR at X band, surface water dynamics via Overhauser dynamic nuclear polarization at X band and short-range distances via cw EPR at high fields.

Milliwatt three- and four-pulse double electron electron resonance for protein structure determination.

Electron paramagnetic resonance (EPR) experiments for protein structure determination using double electron-electron resonance (DEER) spectroscopy rely on high-power microwave amplifiers (>300 W) to create the short pulse lengths needed to excite a sizable portion of the spectrum. The recently introduced self-resonant microhelix combines a high conversion efficiency with an intrinsically large bandwidth (low -value) and a high absolute sensitivity. We report dead times in 3-pulse DEER experiments as low as 14 ± 2 ns achieved using less than 1 W of power at X-band (nominally 9.

Orthogonal spin labeling and pulsed dipolar spectroscopy for protein studies.

Different types of spin labels are currently available for structural studies of biomolecules both in vitro and in cells using Electron Paramagnetic Resonance (EPR) and pulse dipolar spectroscopy (PDS). Each type of label has its own advantages and disadvantages, that will be addressed in this chapter. The spectroscopically distinct properties of the labels have fostered new applications of PDS aimed to simultaneously extract multiple inter-label distances on the same sample.

Strategies to identify and suppress crosstalk signals in double electron-electron resonance (DEER) experiments with gadolinium and nitroxide spin-labeled compounds.

Double electron-electron resonance (DEER) spectroscopy applied to orthogonally spin-labeled biomolecular complexes simplifies the assignment of intra- and intermolecular distances, thereby increasing the information content per sample. In fact, various spin labels can be addressed independently in DEER experiments due to spectroscopically nonoverlapping central transitions, distinct relaxation times, and/or transition moments; hence, they are referred to as spectroscopically orthogonal. Molecular complexes which are, for example, orthogonally spin-labeled with nitroxide (NO) and gadolinium (Gd) labels give access to three distinct DEER channels that are optimized to selectively probe NO-NO, NO-Gd, and Gd-Gd distances.

Biophysical Characterization of Pro-apoptotic BimBH3 Peptides Reveals an Unexpected Capacity for Self-Association.

Bcl-2 proteins orchestrate the mitochondrial pathway of apoptosis, pivotal for cell death. Yet, the structural details of the conformational changes of pro- and antiapoptotic proteins and their interactions remain unclear. Pulse dipolar spectroscopy (double electron-electron resonance [DEER], also known as PELDOR) in combination with spin-labeled apoptotic Bcl-2 proteins unveils conformational changes and interactions of each protein player via detection of intra- and inter-protein distances.

A new perspective on membrane-embedded Bax oligomers using DEER and bioresistant orthogonal spin labels.

Bax is a Bcl-2 protein crucial for apoptosis initiation and execution, whose active conformation is only partially understood. Dipolar EPR spectroscopy has proven to be a valuable tool to determine coarse-grained models of membrane-embedded Bcl-2 proteins. Here we show how the combination of spectroscopically distinguishable nitroxide and gadolinium spin labels and Double Electron-Electron Resonance can help to gain new insights into the quaternary structure of active, membrane-embedded Bax oligomers.

Improved signal fidelity in 4-pulse DEER with Gaussian pulses.

The introduction of arbitrary waveform generator (AWG) technology and the availability of high power microwave amplifiers mark a "new era" in pulse EPR due to significant sensitivity improvements and the possibility to perform novel types of experiments. We present an optimized 4-pulse DEER setup that uses Gaussian observer pulses (GaussDEER) in connection with a Gaussian/shaped pump pulse. Gaussian pulses allow to experimentally remove the "2+1" pulse train ESE signal which is intrinsically present in any DEER experiment performed with rectangular pulses.