A5053413034- X-ray Diffraction in Crystallography
- Enzyme Structure and Function
- Nanoplatforms for cancer theranostics
- Computational Drug Discovery Methods
- Machine Learning in Materials Science
- Inorganic Fluorides and Related Compounds
- Mass Spectrometry Techniques and Applications
- Chemical Thermodynamics and Molecular Structure
- thermodynamics and calorimetric analyses
- S100 Proteins and Annexins
- Biochemical Acid Research Studies
- Radioactive element chemistry and processing
- Biochemical and Molecular Research
- Nanoparticle-Based Drug Delivery
- Genetic Neurodegenerative Diseases
- Microbial metabolism and enzyme function
- Medical Imaging Techniques and Applications
- Superconducting Materials and Applications
- Particle Accelerators and Free-Electron Lasers
- Protein Structure and Dynamics
- Enzyme Production and Characterization
- Genetics, Aging, and Longevity in Model Organisms
- Boron Compounds in Chemistry
- Spectroscopy Techniques in Biomedical and Chemical Research
- Particle Detector Development and Performance
Diamond Light Source
2015-2024
Research Complex at Harwell
2021-2024
Universidade do Vale do Itajaí
2021
University of Oxford
2018
University of Southern Denmark
2011
Brazilian Synchrotron Light Laboratory
2000-2007
Universidade de São Paulo
2004
Abstract COVID-19, caused by SARS-CoV-2, lacks effective therapeutics. Additionally, no antiviral drugs or vaccines were developed against the closely related coronavirus, SARS-CoV-1 MERS-CoV, despite previous zoonotic outbreaks. To identify starting points for such therapeutics, we performed a large-scale screen of electrophile and non-covalent fragments through combined mass spectrometry X-ray approach SARS-CoV-2 main protease, one two cysteine viral proteases essential replication. Our...
Abstract There is currently a lack of effective drugs to treat people infected with SARS-CoV-2, the cause global COVID-19 pandemic. The SARS-CoV-2 Non-structural protein 13 (NSP13) has been identified as target for anti-virals due its high sequence conservation and essential role in viral replication. Structural analysis reveals two “druggable” pockets on NSP13 that are among most conserved sites entire proteome. Here we present crystal structures solved APO form presence both phosphate...
Allostery is an inherent feature of proteins, but it remains challenging to reveal the mechanisms by which allosteric signals propagate. A clearer understanding this intrinsic circuitry would afford new opportunities modulate protein function. Here, we have identified sites in tyrosine phosphatase 1B (PTP1B) combining multiple-temperature X-ray crystallography experiments and structure determination from hundreds individual small-molecule fragment soaks. New modeling approaches 'hidden'...
The severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) macrodomain within the nonstructural protein 3 counteracts host-mediated antiviral adenosine diphosphate-ribosylation signaling. This enzyme is a promising target because catalytic mutations render viruses nonpathogenic. Here, we report massive crystallographic screening and computational docking effort, identifying new chemical matter primarily targeting active site of macrodomain. Crystallographic 2533 diverse fragments...
XChemExplorer (XCE) is a data-management and workflow tool to support large-scale simultaneous analysis of protein-ligand complexes during structure-based ligand discovery (SBLD). The user interfaces established crystallographic software packages such as CCP4 [Winn et al. (2011), Acta Cryst. D67, 235-242] or PHENIX [Adams (2010), D66, 213-221] have entrenched the paradigm that `project' concerned with solving one structure. This does not hold for SBLD, where many almost identical structures...
The steady expansion in the capacity of modern beamlines for high-throughput data collection, enabled by increasing X-ray brightness, robotics and detector speeds, has pushed bottleneck upstream towards sample preparation. Even ligand-binding studies using crystal soaking, experiment best able to exploit beamline capacity, a primary limitation is need gentle nontrivial soaking regimens such as stepwise concentration increases, even robust well characterized crystals. Here, use acoustic...
In fragment-based drug discovery, hundreds or often thousands of compounds smaller than ~300 Da are tested against the protein interest to identify chemical entities that can be developed into potent candidates. Since small, interactions weak, and screening method must therefore highly sensitive; moreover, structural information tends crucial for elaborating these hits lead-like compounds. Therefore, crystallography has always been a gold-standard technique, yet historically too challenging...
Abstract Schistosomiasis is caused by parasites of the genus Schistosoma , which infect more than 200 million people. Praziquantel (PZQ) has been main drug for controlling schistosomiasis over four decades, but despite that it ineffective against juvenile worms and size taste issues with its pharmaceutical forms impose challenges treating school-aged children. It also important to note PZQ resistant strains can be generated in laboratory conditions observed field, hence extensive use mass...
Abstract Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) is the causative agent of disease 2019 (COVID-19). The NSP15 endoribonuclease enzyme, known as NendoU, highly conserved and plays a critical role in ability virus to evade immune system. NendoU promising target for development new antiviral drugs. However, complexity enzyme's structure kinetics, along with broad range recognition sequences lack structural complexes, hampers inhibitors. Here, we performed enzymatic...
PIF1 is a conserved helicase and G4 DNA binding unwinding enzyme, with roles in genome stability. Human (hPIF1) poorly understood, but its functions can become critical for tumour cell survival during oncogene-driven replication stress. Here we report the discovery, via an X-ray crystallographic fragment screen (XChem), of hPIF1 inhibitors. A structure was obtained 4-phenylthiazol-2-amine bound pocket between domains 2A 2B, additional contacts to Valine 258 from domain 1A. The compound makes...
The relationship between GTP binding and hydrolysis on the one hand septin filament assembly function other has yet to be fully elucidated. crystal structures of SEPT3 subgroup members bound either GDP or a analogue shed light this question by indicating how squeezing inter-subunit interfaces may lead exposure polybasic region important for membrane association.
Extracellular exo-inulinase has been isolated from a solid-phase culture of the filamentous fungus Aspergillus awamori var. 2250. The apparent molecular mass monomer enzyme was 69±1kDa, with pI 4.4 and pH optimum 4.5. hydrolysed β-(2 → 1)-fructan (inulin) 6)-fructan (levan) via exo-cleavage, releasing fructose. values for Michaelis constants Km Vmax in hydrolysis inulin were 0.003±0.0001mM 175±5μmol·min−1·mg−1. same parameters levan 2.08±0.04mg/ml 1.2±0.02μmol/min per mg, respectively. gene...
Nicotinic acetylcholine receptors (nAChRs) belong to the family of pentameric ligand-gated ion channels and mediate fast excitatory transmission in central peripheral nervous systems. Among different existing receptor subtypes, homomeric α7 nAChR has attracted considerable attention because its possible implication several neurological psychiatric disorders, including cognitive decline associated with Alzheimer's disease or schizophrenia. Allosteric modulators are particular interest as...
A cowpea class I chitinase (VuChiI) was expressed in the methylotrophic yeast P. pastoris. The recombinant protein secreted into culture medium and purified by affinity chromatography on a chitin matrix. migrated SDS-polyacrylamide gel electrophoresis as two closely-related bands with apparent molecular masses of 34 37 kDa. identity these VuChiI demonstrated mass spectrometry analysis tryptic peptides N-terminal amino acid sequencing. able to hydrolyze colloidal but did not exhibit enzymatic...
ABSTRACT Fragment‐based drug discovery is a well‐established method for the identification of chemical starting points development into clinical candidates. Historically, crystallographic fragment screening was perceived to be low‐throughput and time consuming. However, thanks advances in synchrotron capabilities introduction dedicated facilities, such as XChem platform at Diamond Light Source, there have been substantial improvements throughput integration between sample preparation, data...
Citrus canker, caused by bacteria Xanthomonas citri subsp. citri, can affect all economically important varieties of citrus. Studying genes related to the invasive capacity may improve knowledge on how this works and ultimately use information avoid disease. Some annotated from published genome are addressed an interesting class named "pathogenicity, virulence adaptation". One them is xanA, which encodes a predicted phosphoglucomutase. Phosphoglucomutases ubiquitous enzymes among living...
Extracellular exo-inulinase has been isolated from a solid-phase culture of the filamentous fungus Aspergillus awamori var. 2250. The apparent molecular mass monomer enzyme was 69±1kDa, with pI 4.4 and pH optimum 4.5. hydrolysed β-(2 → 1)-fructan (inulin) 6)-fructan (levan) via exo-cleavage, releasing fructose. values for Michaelis constants Km Vmax in hydrolysis inulin were 0.003±0.0001mM 175±5μmol·min−1·mg−1. same parameters levan 2.08±0.04mg/ml 1.2±0.02μmol/min per mg, respectively. gene...