Biophysical Society 66th Annual Meeting Program Guide

3:30 PM – 5:00 PM Dynamic Biosensors Measurement of PROTAC Ternary Complex Formation Using the switchSENSE® Y-Structure and FRET Signals Proteolysis targeting chimeras (PROTACs) are essential bifunctional small molecules that engage the formation of a ternary complex consist- ing of an E3 ubiquitin ligase, a target protein of interest and the PROTAC itself. Using switchSENSE® technology and the novel DNA Y-structure, an E3 ligase as well as a target protein can be functionalized on each separate end of two FRET pair color-coded Y-arms. The Y-structure closes upon PROTAC binding and the subsequent ternary complex formation bringing together the green donor and the red acceptor dye into a closer, FRET sensitive, distance. The change in red fluorescence signal intensity directly correlates with ternary complex formation kinetics. Here, we show that the Y-structure is an extremely versatile tool for studying any type of protein-protein complex formations with a dis- sociation constant between 1nM to 10µM. With the switchSENSE® technology and the highly sensitive FRET read-out, it is possible to perform high-throughput PROTAC screening and to characterize their kinetics (PROTACs ranking), gaining information on binary and ternary binding at the same time. Speaker Jonathan Faherty, Head of Operations, Dynamic Biosensors Esplanade, Room 157: Monday, February 21 9:30 AM – 11:00 AM Bruker Recent Advancements in Magnetic Resonance Solutions for Biology Magnetic Resonance offers many insights into how biological sys- tems function. Nuclear Magnetic Resonance (NMR) and Electron Paramagnetic Resonance (EPR) are two techniques used to identity species, dynamics, and structures of proteins, peptides, nucleotides, and lipids. The speakers in this session will present recent advance- ments in magnetic resonance in biophysics. Clemens Anklin will present “Optimizing the NMR Workflow for Protein Dynamics Experiments." NMR is ideally suited to study proteins in motion. A suite of NMR experiments allows the ability to look at pro- tein dynamics from picosecond to very long timescales. One of the difficulties in performing these experiments was the need to assemble and master a variety of tools. We are currently working on creating an integrated workflow that can facilitate this task for the researcher. From experiment selection, to import of assignments, to analysis of the experimental data to visualization on a structure the user can find all these tools in a single workflow. Ralph Weber will present “New EPR Tools for Studying Biological Structures and Dynamics” Motional dynamics in biological molecules such as proteins, peptides, and nucleotides, offers great insights into the functioning of these biological systems. Owing to the broad range of time scales accessible to EPR experiments, coupled with its high sensitivity compared to other techniques, EPR has been invaluable in advancing knowledge of many biological processes. Analysis of line shapes yields the desired information. Bruker now offers the SpinFit

Liquids software module to easily extract information regarding the motional dynamics. Fitting of spectra is fully implemented and mul- tiple species can be analyzed simultaneously. The module is compat- ible with all current Bruker spectrometers. DEER (Double Electron Electron Resonance) has proven to be very successful in structural biology studies. Each sample is pre-screened to evaluate the labelling efficiency. With the introduction of the Bruker Magnettech ESR5000, we now offer a reasonably priced benchtop option (about the size of a UV/Vis spectrometer) that has the sensitivity to acquire the spectra and also is available with the optional SpinFit Liquids and SpinCount modules to ascertain the labelling efficiency. Screening is performed off-line, thus not interfer- ing with the on-going DEER experiments to increase lab productivity. In addition, the spectrometer helps in pre-screening DNP samples. EPR accessibility measurements are a valuable molecular ruler for peptides and proteins in membranes for structural studies. The Bruker Magnettech ESR5000 enables this important experimental technique. Rapid Scan EPR offers improvements in sensitivity. We invite you to see the latest results applying the technique to biophysical studies. Speakers Clemens Anklin, Vice President of NMR Applications, Bruker Biospin Ralph Weber, Senior Applications Scientist, Bruker Biospin 11:30 AM – 1:00 PM LEICA MICROSYSTEMS INC Label-Free, Chemically Specific Imaging with the Leica STELLARIS 8 CRS Coherent Raman Scattering microscopy (CRS) is a powerful new imaging technique that provides label-free, chemically specific image contrast based on the characteristic intrinsic vibrational of the sam- ple molecules. CRS can produce high-resolution (sub-cellular level), dynamic (up to video-rate), and quantifiable information on the bio- chemical composition and metabolic processes in cells, tissues, and intact model organisms, and it enables imaging of small molecules without perturbing their function. This information is highly synergis- tic with the types of contrast provided by fluorescence microscopy. Here, I will provide an overview of the wide range of application areas covered by the all-new STELLARIS 8 CRS – Leica’s hands-free Coherent Raman Scattering microscope. The instrument offers both CRS modalities – Stimulated Raman Scattering (SRS) and CARS – and allows for the simultaneous acquisition of two-photon fluorescence and second-harmonic generation signals. Importantly, the seamless integration of CRS with the STELLARIS visible confocal fluorescence microscopy platform results in a true multi-modal optical discovery instrument that is capable of capturing a unique combination of biochemical, biophysical and molecular contrasts. Unsurprisingly, the instrument is finding a growing number of applications in fields like neurodegenerative disease, cancer, 3D biology, stem cell and devel- opmental biology, and pharmacology. Speaker Volker Schweikhard, Global Application Manager, LEICA MICROSYSTEMS INC

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