A5084479435- Hemoglobin structure and function
- Erythrocyte Function and Pathophysiology
- Protein Structure and Dynamics
- Electron Spin Resonance Studies
- Pharmacogenetics and Drug Metabolism
- Neonatal Health and Biochemistry
- Trypanosoma species research and implications
- Research on Leishmaniasis Studies
- Advanced NMR Techniques and Applications
- Enzyme-mediated dye degradation
- Quantum Electrodynamics and Casimir Effect
- SARS-CoV-2 and COVID-19 Research
- Immunotherapy and Immune Responses
- Heme Oxygenase-1 and Carbon Monoxide
- vaccines and immunoinformatics approaches
- Biofuel production and bioconversion
- Calcium signaling and nucleotide metabolism
- Polysaccharides and Plant Cell Walls
- Fungal and yeast genetics research
- Enzyme function and inhibition
- Amyotrophic Lateral Sclerosis Research
- Porphyrin Metabolism and Disorders
- Bacteriophages and microbial interactions
- Synthesis and Catalytic Reactions
- Bacterial Genetics and Biotechnology
Institute of Molecular Biology and Pathology
2021-2025
National Research Council
2022-2024
Sapienza University of Rome
2018-2024
Istituto Pasteur
2019-2020
Trypanothione reductase (TR) is a suitable target for drug discovery approaches against leishmaniasis, although the identification of potent inhibitors still challenging. Herein, we harnessed fragment-based (FBDD) strategy to develop new TR inhibitors. Previous crystallographic screening identified fragments
COVID-19 is a highly infectious disease caused by newly emerged coronavirus (SARS-CoV-2) that has rapidly progressed into pandemic. This unprecedent emergency stressed the significance of developing effective therapeutics to fight current and future outbreaks. The receptor-binding domain (RBD) SARS-CoV-2 surface Spike protein main target for vaccines represents helpful "tool" produce neutralizing antibodies or diagnostic kits. In this work, we provide detailed characterization native RBD...
The regulation of cytochrome P450 activity is often achieved by structural transitions induced substrate binding. We describe the conformational transition experienced upon binding OleP, an epoxygenase involved in oleandomycin biosynthesis. OleP bound to analog 6DEB crystallized 2 forms: one with ensemble open and closed conformations asymmetric unit another only conformation. Characterization O1eP-6DEB kinetics, also using inhibitor clotrimazole, unveiled a complex mechanism that involves...
Trypanothione reductase (TR) is a key factor in the redox homeostasis of trypanosomatid parasites, critical for survival hostile oxidative environment generated by host to fight infection. TR considered an attractive target development new trypanocidal agents as it essential parasite but has no close homolog humans. However, high efficiency and turnover challenging targets since only potent inhibitors, with nanomolar IC50, can significantly affect state viability. To aid design effective...
Different murine neuroglobin variants showing structural and dynamic alterations that are associated with perturbation of ligand binding have been studied: the CD loop mutants characterized by an enhanced flexibility (Gly-loop40-48 Gly-loop44-47 ), F106A mutant, double /F106A mutant. Their ferric resonance Raman spectra in solution crystals almost identical. In high-frequency region, identification a set core size marker bands indicates presence two 6-coordinate low spin species. The data,...
Background: As a result of the paucity treatment, Leishmaniasis continues to provoke about 60,000 deaths every year worldwide. New molecules are needed, and drug discovery research is oriented toward targeting proteins crucial for parasite survival. Among them, trypanothione reductase (TR) remarkable interest owing its vital role in Leishmania species protozoan life. Our previously identified compound 1 novel chemotype endowed with unique mode TR inhibition thanks binding formerly unknown...
Abstract Neuroglobin (Ngb) is predominantly expressed in neurons of the central and peripheral nervous systems it clearly seems to be involved neuroprotection. Engineering Ngb observe structural dynamic alterations associated with perturbation ligand binding might reveal important determinants, could shed light on key features related its mechanism action. Our results highlight relevance CD loop Phe106 as distal proximal controls murine neuroglobin. We observed effects individual combined...
Substrate binding to the cytochrome P450 OleP is coupled a large open-to-closed transition that remodels active site, minimizing its exposure external solvent. When aglycone substrate binds, small empty cavity formed between I and G helices, BC loop, itself, where solvent molecules accumulate mediating substrate-enzyme interactions. Herein, we analyzed role of this in by producing three mutants (E89Y, G92W, S240Y) decrease volume. The crystal structures closed state bound 6DEB showed G92W...
Abstract Background Fungal glucose dehydrogenases (GDHs) are FAD-dependent enzymes belonging to the glucose-methanol-choline oxidoreductase superfamily. These classified in “Auxiliary Activity” family 3 (AA3) of Carbohydrate-Active enZymes database, and more specifically subfamily AA3_2, that also includes closely related flavoenzymes aryl-alcohol oxidase 1-oxidase. Based on sequence similarity known fungal GDHs, an AA3_2 enzyme active was identified genome Pycnoporus cinnabarinus , a model...
Leishmania spp. are responsible for up to 1 million new cases each year. The current therapeutic arsenal against is largely inadequate, and there an urgent need better drugs. Trypanothione reductase (TR) represents a druggable target since it essential the parasite not shared by human host. Here, we report optimization of novel class potent selective LiTR inhibitors realized through concerted effort involving X-ray crystallography, synthesis, structure–activity relationship (SAR)...
A combined biophysical approach was applied to map gas-docking sites within murine neuroglobin (Ngb), revealing snapshots of events that might govern activity and dynamics in this unique hexacoordinate globin, which is most likely be involved gas-sensing the central nervous system for a precise mechanism action remains elucidated. The application UV–visible microspectroscopy crystallo , solution X-ray absorption near-edge spectroscopy diffraction experiments at 15–40 K provided structural...
The cytochrome P450 OleP catalyzes the epoxidation of aliphatic carbons on both aglycone 8.8a-deoxyoleandolide (DEO) and monoglycosylated L-olivosyl-8.8a-deoxyoleandolide (L-O-DEO) intermediates oleandomycin biosynthesis. We investigated substrate versatility enzyme. X-ray equilibrium binding data show that DEO loosely fits active site, triggering closure prepares it for catalysis only a minor population open-to-closed state transition allows solvent molecules to accumulate in cavity forms...
Specificity in protein-DNA recognition arises from the synergy of several factors that stem structural and chemical signatures encoded within targeted DNA molecule. Here, we deciphered nature interactions driving binding by bacterial transcription factor PdxR, a member MocR family responsible for regulation pyridoxal 5'-phosphate (PLP) biosynthesis. Single particle cryo-EM performed on PLP-PdxR bound to its target enabled isolation three conformers complex, which may be considered as...
Abstract In heme proteins, the canonical and reversed conformations result from rotation of group by 180° about α,γ‐meso axis in protein pocket. The coexistence two different orientations has been observed both proteins reconstituted with hemin some native proteins. reversal orientation can also change certain functional properties Complementing results other experimental techniques, like circular dichroism nuclear magnetic resonance, resonance Raman spectroscopy provides detailed...
We produced a neuroglobin variant, namely, Ngb CDless, with the excised CDloop- and D-helix, directly joining C- E-helices. The CDless variant retained bis-His hexacoordination, we investigated role of CDloop–D-helix unit in controlling CO binding structural dynamics by an integrative approach based on X-ray crystallography, rapid mixing, laser flash photolysis, resonance Raman spectroscopy, molecular simulations. Rapid mixing photolysis showed that ligand affinity was unchanged respect to...
Abstract COVID-19 is a highly infectious disease caused by newly emerged coronavirus (SARS-CoV-2) that has rapidly progressed into pandemic. This unprecedent emergency stressed the significance of developing effective therapeutics to fight current and future outbreaks. The receptor-binding domain (RBD) SARS-CoV-2 surface Spike protein main target for vaccines represents helpful “tool” produce neutralizing antibodies or diagnostic kits. In this work, we provide detailed characterization...
Abstract Trypanothione reductase from Trypanosoma brucei ( Tb TR) is a key enzyme of the parasite responsible for human African trypanosomiasis. TR catalyzes reduction trypanothione, an antioxidant dithiol that protects trypanosomatid parasites oxidative stress induced by mammalian host as defense system. considered attractive target development novel anti‐parasitic drugs because it essential survival and has no close homologues in humans. Previous studies have shown metal compounds can act...
Cytochrome P450 OleP catalytic activity is strongly influenced by its structural dynamic conformational behavior. Here, we combine equilibrium-binding experiments with all-atom molecular dynamics simulations to clarify how different environments affect equilibrium between the open and closed-catalytic competent-forms. Our data clearly show that at high-ionic strength conditions, closed form favored, and, very interestingly, mechanisms, depending on chemistry of cations, can be used...