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he Beam Dump Experiment (BDX) at Jefferson Laboratory (JLab) is an electron-beam thick-target experiment to search for Light Dark Matter (LDM) particles in the MeV-GeV mass range. BDX will exploit the high-intensity 10.6 GeV e− beam from CEBAF accelerator impinging on the beam dump of experimental Hall-A, collecting up to 1022 electrons-on-target (EOT) in a few years time. Any LDM particle produced by the interaction of the primary e− beam with the beam dump will be detected by measuring their scattering inside the BDX detector, an electromagnetic calorimeter surrounded by an hermetic veto system, which is to be installed in a dedicated underground facility, located 20 m downstream. Thanks to the large detection efficiency and background rejection capabilities, BDX will be able to explore a so-far unknown region in the LDM parameter space, improving current exclusion limits by one order of magnitude in case of a null observation. 

The BGOOD photoproduction experiment accesses forward meson angles and low momentum exchange kinematics in the uds sector, which may be sensitive to molecular-like hadronic structure. Recent highlights are summarised in these proceedings. 

Protein labeling strategies have been explored for decades to study protein structure, function, and regulation. Fluorescent labeling of a protein enables the study of protein‐protein interactions through biophysical methods such as microscale thermophoresis (MST). MST measures the directed motion of a fluorescently labeled protein in response to microscopic temperature gradients, and the protein's thermal mobility can be used to determine binding affinity. However, the stoichiometry and site specificity of fluorescent labeling are hard to control, and heterogeneous labeling can generate inaccuracies in binding measurements. Here, we describe an easy‐to‐apply protocol for high‐stoichiometric, site‐specific labeling of a protein at its N‐terminus withN‐hydroxysuccinimide (NHS) esters as a means to measure protein‐protein interaction affinity by MST. This protocol includes guidelines for NHS ester labeling, fluorescent‐labeled protein purification, and MST measurement using a labeled protein. As an example of the entire workflow, we additionally provide a protocol for labeling a ubiquitin E3 enzyme and testing ubiquitin E2‐E3 enzyme binding affinity. These methods are highly adaptable and can be extended for protein interaction studies in various biological and biochemical circumstances. © 2021 Wiley Periodicals LLC.

This article was corrected on 18 July 2022. See the end of the full text for details.

Basic Protocol 1: Labeling a protein of interest at its N‐terminus with NHS esters through stepwise reaction

Alternate Protocol: Labeling a protein of interest at its N‐terminus with NHS esters through a one‐pot reaction

Basic Protocol 2: Purifying the N‐terminal fluorescent‐labeled protein and determining its concentration and labeling efficiency

Basic Protocol 3: Using MST to determine the binding affinity of an N‐terminal fluorescent‐labeled protein to a binding partner.

Basic Protocol 4: NHS ester labeling of ubiquitin E3 ligase WWP2 and measurement of the binding affinity between WWP2 and an E2 conjugating enzyme by the MST binding assay

 

The BGO-OD experiment at the ELSA accelerator facility uses an energy tagged bremsstrahlung photon beam to investigate the excitation structure of the nucleon. The setup consists of a highly segmented BGO calorimeter surrounding the target, with a particle tracking magnetic spectrometer at forward angles. BGO-OD is ideal for investigating low momentum transfer processes due to the acceptance and high momentum resolution at forward angles. In particular, this enables the investigation of strangeness photoproduction where t-channel exchange mechanisms play an important role. This also allows access to low momentum exchange kinematics where extended, molecular structure may manifest in reaction mechanisms. First key results at low t indicate a cusp-like structure in K + Σ 0 photoproduction at W = 1900 MeV, line shapes and differential cross sections for K + Λ(1405)→ K + Σ 0 π 0 , and a peak structure in K 0 S Σ 0 photoproduction. The peak in the K 0 S Σ 0 channel appears consistent with meson-baryon generated states, where equivalent models have been used to describe the P C pentaquark candidates in the heavy charmed quark sector. 

Since the discovery of the Λ(1405), it remains poorly described by conventional constituent quark models, and it is a candidate for having an “exotic” meson-baryon or “penta-quark” structure, similar to states recently reported in the hidden charm sector. The Λ(1405) can be produced in the reaction γp K + Λ(1405). The pure I=0 decay mode into Σ 0 π 0 is prohibited for the mass-overlapping Σ(1385). Combining a large aperture forward magnetic spectrometer and a central BGO crystal calorimeter, the BGO-OD experiment is ideally suited to measure this decay with the K + in the forward direction. Preliminary results are presented. *Supported by DFG (PN 388979758, 405882627) 

The type I insulin-like growth factor receptor (IGF1R) is involved in growth and survival of normal and neoplastic cells. A ligand-dependent conformational change is thought to regulate IGF1R activity, but the nature of this change is unclear. We point out an underappreciated dimer in the crystal structure of the related Insulin Receptor (IR) with Insulin bound that allows direct comparison with unliganded IR and suggests a mechanism by which ligand regulates IR/IGF1R activity. We test this mechanism in a series of biochemical and biophysical assays and find the IGF1R ectodomain maintains an autoinhibited state in which the TMs are held apart. Ligand binding releases this constraint, allowing TM association and unleashing an intrinsic propensity of the intracellular regions to autophosphorylate. Enzymatic studies of full-length and kinase-containing fragments show phosphorylated IGF1R is fully active independent of ligand and the extracellular-TM regions. The key step triggered by ligand binding is thus autophosphorylation.