Spin-label scanning reveals conformational sensitivity of the bound helical interfaces of IA₃

IA3 is an intrinsically disordered protein (IDP) that becomes helical when bound to yeast proteinase A (YPRA) or in the presence of the secondary stabilizer 2,2,2-trifluoroethanol (TFE). Here, site-directed spin-labeling (SDSL) continuous wave electron paramagnetic resonance (CW-EPR) spectroscopy and circular dichroism (CD) are used to characterize the TFE-induced helical conformation of IA3 for a series of spin-labeled cysteine scanning constructs and varied amino acid substitutions. Results demonstrate that the N-terminal concave helical surface of IA3, which is the buried interface when bound to YPRA, can be destabilized by the spin-label or bulky amino acid substitutions. In contrast, the helical tendency of IA3 is enhanced when spin-labels are incorporated into the convex, i.e., solvent exposed, surface of IA3. No impact of the spin-label within the C-terminal region was observed. This work further demonstrates the utility and sensitivity of SDSL CW-EPR for studies of IDPs. In general, care must be taken to ensure the spin-label does not interfere with native helical tendencies and these studies provide us with knowledge of where to incorporate spin-labels for future SDSL investigations of IA3

Effects of PRE and POST therapy drug-pressure selected mutations on HIV-1 protease conformational sampling

AbstractConformational sampling of pre- and post-therapy subtype B HIV-1 protease sequences derived from a pediatric subject infected via maternal transmission with HIV-1 were characterized by double electron–electron resonance spectroscopy. The conformational ensemble of the PRE construct resembles native-like inhibitor bound states. In contrast, the POST construct, which contains accumulated drug-pressure selected mutations, has a predominantly semi-open conformational ensemble, with increased populations of open-like states. The single point mutant L63P, which is contained in PRE and POST, has decreased dynamics, particularly in the flap region, and also displays a closed-like conformation of inhibitor-bound states. These findings support our hypothesis that secondary mutations accumulate in HIV-1 protease to shift conformational sampling to stabilize open-like conformations, while maintaining the predominant semi-open conformation for activity

Molecular profiling reveals immunogenic cues in anaplastic large cell lymphomas with DUSP22 rearrangements

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Studying the composition and mineralogy of the hermean surface with the Mercury Radiometer and Thermal Infrared Spectrometer (MERTIS) for the BepiColombo mission: an update

Launched onboard the BepiColombo Mercury Planetary Orbiter (MPO) in October 2018, the Mercury Radiometer and Thermal Infrared Spectrometer (MERTIS) is on its way to planet Mercury. MERTIS consists of a push-broom IR-spectrometer (TIS) and a radiometer (TIR), which operate in the wavelength regions of 7-14 μm and 7-40 μm, respectively. This wavelength region is characterized by several diagnostic spectral signatures: the Christiansen feature (CF), Reststrahlen bands (RB), and the Transparency feature (TF), which will allow us to identify and map rock-forming silicates, sulfides as well as other minerals. Thus, the instrument is particularly well-suited to study the mineralogy and composition of the hermean surface at a spatial resolution of about 500 m globally and better than 500 m for approximately 5-10% of the surface. The instrument is fully functional onboard the BepiColombo spacecraft and exceeds all requirements (e.g., mass, power, performance). To prepare for the science phase at Mercury, the team developed an innovative operations plan to maximize the scientific output while at the same time saving spacecraft resources (e.g., data downlink). The upcoming fly-bys will be excellent opportunities to further test and adapt our software and operational procedures. In summary, the team is undertaking action at multiple levels, including performing a comprehensive suite of spectroscopic measurements in our laboratories on relevant analog materials, performing extensive spectral modeling, examining space weathering effects, and modeling the thermal behavior of the hermean surface