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Cryo-EM structure of AAV2 Rep68 bound to integration site AAVS1: insights into the mechanism of DNA melting.
Nucleic Acids Res
January 2025
Department of Physiology and Biophysics, Virginia Commonwealth University, School of Medicine, Richmond, VA 23298, United States.
The Rep68 protein from Adeno-Associated Virus (AAV) is a multifunctional SF3 helicase that performs most of the DNA transactions necessary for the viral life cycle. During AAV DNA replication, Rep68 assembles at the origin of replication, catalyzing the DNA melting and nicking reactions during the hairpin rolling replication process to complete the second-strand synthesis of the AAV genome. We report the cryo-electron microscopy structures of Rep68 bound to the adeno-associated virus integration site 1 in different nucleotide-bound states.
View Article and Find Full Text PDFMultiplex basophil activation tests for allergy diagnosis: present and future applications.
Front Allergy
January 2025
Laboratory for Clinical Immunology and Molecular Genetics, University Clinic of Respiratory and Allergic Diseases Golnik, Golnik, Slovenia.
The basophil activation test (BAT) has become a major cellular test for evaluating the allergenic activity of specific IgEs. The impact of the BAT is due to the ability of blood basophil granulocytes to present IgE on the high-affinity FcRI receptor and to mirror the mast cell response that elicits an acute allergic reaction. The BAT proved to be able to identify allergic patients at risk of reacting to a low dose of the allergen and/or developing life-threatening reactions and thus can significantly improve the current management of allergic patients.
View Article and Find Full Text PDFGlycan-Matchmade Multivalent Decoration of Enzyme Labels for Amplified Electrochemical Detection of Glycoproteins.
Anal Chem
January 2025
Center for Advanced Analytical Science, Guangzhou Key Laboratory of Sensing Materials and Devices, Guangdong Engineering Technology Research Center for Sensing Materials and Devices, School of Chemistry and Chemical Engineering, Guangzhou University, Guangzhou 510006, China.
Glycoproteins are of significant value to liquid biopsy of human diseases. Herein, we present a universal electrochemical platform for the amplified detection of glycoproteins, taking advantage of the glycan-matchmade multivalent decoration of enzyme labels for the enzymatic signal amplification. Briefly, the glycan-matchmade multivalent decoration involves two steps, i.
View Article and Find Full Text PDFA suite of pre-assembled, pET28b-based Golden Gate vectors for efficient protein engineering and expression.
bioRxiv
January 2025
Department of Cell Biology, University of Pittsburgh, Pittsburgh, PA 15261.
Expression and purification of recombinant proteins in is a bedrock technique in biochemistry and molecular biology. Expression optimization requires testing different combinations of solubility tags, affinity purification techniques, and site-specific proteases. This optimization is laborious and time consuming as these features are spread across different vector series and require different cloning strategies with varying efficiencies.
View Article and Find Full Text PDFUncovering the Mechanism of Action of Antiprotozoal Agents: A Survey on Photoaffinity Labeling Strategy.
Pharmaceuticals (Basel)
December 2024
Dipartimento di Scienze della Vita, della Salute e delle Professioni Sanitarie, Università degli Studi "Link Campus University", Via del Casale di S. Pio V 44, I-00165 Rome, Italy.
, , and parasites are responsible for infectious diseases threatening millions of people worldwide. Despite more recent efforts devoted to the search for new antiprotozoal agents, efficacy, safety, and resistance issues still hinder the development of suited therapeutic options. The lack of robustly validated targets and the complexity of parasite's diseases have made phenotypic screening a preferential drug discovery strategy for the identification of new chemical entities.
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