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Light-Induced Conformational Heterogeneity Induces Positive Photoswitching in Photoconvertible Fluorescent Proteins of the EosFP Family.
J Am Chem Soc
March 2025
Université Grenoble Alpes, CEA, CNRS, Institut de Biologie Structurale (IBS), 71 Avenue des Martyrs, Grenoble, Cedex 9 38044, France.
Green-to-red photoconvertible fluorescent proteins (PCFPs) of the EosFP family are commonly used in ensemble pulse-chase and single-molecule localization or tracking approaches. However, these fluorescent proteins exhibit highly complex photophysical behaviors. In the green-form, recent NMR experiments revealed that mEos4b and other PCFP variants exist in two different conformational states at thermal equilibrium, which limits their effective photoconversion efficiency.
View Article and Find Full Text PDFMechanism of Conformational Selection of tRNA by Bacterial Deaminase TadA.
Biochemistry
March 2025
Department of Chemistry, Indian Institute of Technology Bombay, Powai, Mumbai 400076, India.
Base editing is a common mechanism by which organisms expand their genetic repertoire to access new functions. Here, we explore the mechanism of tRNA recognition in the bacterial deaminase TadA, which exclusively recognizes tRNA and converts the wobble base adenosine (A34) to inosine. We quantitatively evaluate the dynamics of tRNA binding by incorporating the fluorescent adenine analogue 2-aminopurine (2-AP) at position 34 in the wobble base of the anticodon loop.
View Article and Find Full Text PDFElectroporation Techniques to Target Subependymal Zone Neurogenic Niche.
Methods Mol Biol
March 2025
Instituto de Biotecnología y Biomedicina (BIOTECMED), Departamento de Biología Celular, Universidad de Valencia, Burjassot, Valencia, Spain.
In vivo electroporation is a technique that allows for the transfection of specific neuronal progenitor pools in the developing embryo or during early postnatal stages. By combining plasmid DNA injection with the application of electric pulses, this method directs DNA to the targeted region, facilitating its entry into neural stem cells. This approach enables functional experiments by perturbing genes of interest and conducting cell-tracking studies using fluorescent proteins.
View Article and Find Full Text PDFDuplexed CeTEAM drug biosensors reveal determinants of PARP inhibitor selectivity in cells.
J Biol Chem
February 2025
Science for Life Laboratory, Division of Clinical Physiology, Department of Laboratory Medicine, Karolinska Institutet, Karolinska University Hospital; Huddinge, SE-141 52, Sweden. Electronic address:
Poly (ADP-ribose) polymerase (PARP) inhibitors (PARPi) targeting PARP1 and PARP2 have revolutionized cancer therapy by selectively killing cancer cells with defective DNA repair. However, achieving PARP1 or PARP2-selective inhibitors is difficult due to structural homology. Selectivity profiling is typically done with purified proteins, but these lack the complexity of intracellular environments and could therefore be inaccurate.
View Article and Find Full Text PDFA caged luciferin analogue generating near-infrared bioluminescence for activity-sensing of labile iron.
Biosens Bioelectron
February 2025
Department of Radiology, Zhongnan Hospital of Wuhan University, Wuhan, 430071, PR China. Electronic address:
Iron plays a vital role in physiological processes due to its high oxygen affinity and efficient redox capability. However, perturbations in iron homeostasis, particularly in its labile forms that drive oxidative stress, have been implicated in a spectrum of pathologies, including infectious diseases, malignancies, and neurodegenerative disorders. Despite the critical importance of detecting labile Fe, conventional fluorescent and bioluminescent probes are constrained by inherent limitations, such as suboptimal sensitivity, elevated background noise, and inadequate tissue penetration depth.
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