Docente
Jose Artur Alves De Brito

Resume

Since completing my 5 year-B.Sc. degree in Biotechnological Engineering in 2001, followed by a Ph.D. degree in Biochemistry in 2011, I have gathered a long experience in the field of Structural Biology. My focus has been the 3D structure determination of macromolecules using X-ray crystallography. Recently, I have started also to use cryo-electron microscopy (cryo-EM) to address proteins recalcitrant to crystallization, and further expand my repertoire in structural biology techniques. After a short stay at the University of Madison (Wisconsin, USA), where I gained expertise in protein purification and functional characterization of the human cardiac protein myosin, I pursued my doctoral studies at ITQB NOVA (Oeiras, Portugal), and the University of Leeds (UK). There, I studied enzymes from extremophilic organisms to gain insights into their specific metabolic pathways and the key structural determinants contributing to their remarkable stability. These formative experiences at foreign institutions greatly shaped my current multidisciplinary, collaborative, and result-driven scientific perspective. After completing my Ph.D., I took a joint postdoctoral position with the Membrane Protein Crystallography (MPX) and the Bacterial Cell Biology labs at ITQB NOVA. During this time, I gained experience working with membrane two-component systems of the human pathogen Staphylococcus aureus as putative sensors of antibiotic presence. Simultaneously, I continued to investigate sulfur-metabolizing pathways. In 2018, I assumed a Junior Research position at ITQB NOVA and started to gain autonomy in research lines involving sulfur metabolism, particularly on hydrogen sulfide synthesis and degradation, thiosulfate/tetrathionate interconversion, and RNA homeostasis pathways. At this moment, my research interests became clear and focused: some of the proteins involved in these pathways are from pathogenic organisms and represent potential novel drug targets which can be used to tackle antibiotic resistance, a serious public health threat. Noteworthy, the autonomy I have gained in this scientific position granted me the possibility to develop my research objectives, build a scientific network, supervise B.Sc. and Master students and build my team. In 2019, I became the acting co-Principal Investigator of the MPX Laboratory at ITQB NOVA headed by Dr. Margarida Archer. This position provided me with further experience in training and supervision of Ph.D. students working on many different projects. In addition, I assumed the main responsibilities for the daily management of the MPX Lab, including regular communication with the accounting, academics, human resources, and purchase departments of the Institute. I have also contributed significantly to boosting the development of the cryo-EM capacity at ITQB NOVA, namely in purchase acquisition procedures and establishing/managing cryo-EM data storage and processing facilities at our Institute. Additionally, I was appointed co-manager of the automated crystallization facility at ITQB NOVA. Through this path, I became highly skilled in macromolecular crystallography including protein crystallization, data collection at synchrotrons, data processing and structure determination, model building, and crystallographic refinement. I am also proficient in protein biophysics techniques (Thermal Shift Assay, nano-Differential Scanning Fluorimetry, and Dynamic Light Scattering) along with the biochemical, spectroscopic, and functional characterization of enzymes. I consider myself a good-natured, resilient, and hard-working scientist, with a strong focus on teamwork. I am highly organized and goal-oriented, relying on data to guide my decision-making and solution-driven approach. My ambition and drive are balanced by a positive and enthusiastic personality, and I feel well-prepared to take on new challenges and achieve any outlined objectives, whatever these might be.

Graus

• Licenciatura
  Engenharia Biotecnológica
• Doutoramento
  Bioquímica
• Pós-doutoramento
  Targeting antimicrobial resistance with an X-ray "machine gun": the two-component system VraSR from Staphylococcus aureus

Publicações

Journal article

• 2023-10, Structure and function of Campylobacter jejuni polynucleotide phosphorylase (PNPase): Insights into the role of this RNase in pathogenicity, Biochimie
• 2022-12-15, Urolithin B: Two-way attack on IAPP proteotoxicity with implications for diabetes., Frontiers in endocrinology
• 2022-11, The staphylococcal inhibitory protein SPIN binds to human myeloperoxidase with picomolar affinity but only dampens halide oxidation, Journal of Biological Chemistry
• 2022-08-19, Metabolic and Structural Insights into Hydrogen Sulfide Mis-Regulation in Enterococcus faecalis, Antioxidants
• 2022-05, Biochemical and Biophysical Characterization of Exoribonucleases from the Human Pathogen Mycobacterium tuberculosis, Access Microbiology
• 2020-08, A scalable insect cell-based production process of the human recombinant BMX for in-vitro covalent ligand high-throughput screening., Bioprocess and biosystems engineering
• 2020-03-20, 3-Oxo-ß-sultam as a Sulfonylating Chemotype for Inhibition of Serine Hydrolases and Activity-Based Protein Profiling, ACS Chemical Biology
• 2018-12, X-ray structure of full-length human RuvB-Like 2 – mechanistic insights into coupling between ATP binding and mechanical action, Scientific Reports
• 2017, The key role of glutamate 172 in the mechanism of type II NADH:quinone oxidoreductase of Staphylococcus aureus, Biochimica et Biophysica Acta - Bioenergetics
• 2017, A Clinically Relevant Variant of the Human Hydrogen Sulfide-Synthesizing Enzyme Cystathionine ß -Synthase: Increased CO Reactivity as a Novel Molecular Mechanism of Pathogenicity?, Oxidative Medicine and Cellular Longevity
• 2016, Electron Accepting Units of the Diheme Cytochrome c TsdA, a Bifunctional Thiosulfate Dehydrogenase/Tetrathionate Reductase, Journal of Biological Chemistry
• 2016, Clickable 4-Oxo-ß-lactam-Based Selective Probing for Human Neutrophil Elastase Related Proteomes, ChemMedChem
• 2015, Type-II NADH: Quinone oxidoreductase from Staphylococcus aureus has two distinct binding sites and is rate limited by quinone reduction, Molecular Microbiology
• 2015, Thiosulfate Dehydrogenase (TsdA) from Allochromatium vinosum STRUCTURAL AND FUNCTIONAL INSIGHTS INTO THIOSULFATE OXIDATION, Journal of Biological Chemistry
• 2015, Structural and Functional Characterization of an Ancient Bacterial Transglutaminase Sheds Light on the Minimal Requirements for Protein Cross-Linking, Biochemistry
• 2015, Stabilization of porcine pancreatic elastase crystals by glutaraldehyde cross-linking, Acta Crystallographica Section:F Structural Biology Communications
• 2015, Expression, purification, crystallization and preliminary X-ray diffraction analysis of a type II NADH:quinone oxidoreductase from the human pathogen Staphylococcus aureus, Acta Crystallographica Section F:Structural Biology Communications
• 2014, Production, crystallization and preliminary crystallographic analysis of Allochromatium vinosum thiosulfate dehydrogenase TsdA, an unusual acidophilic c-type cytochrome, Acta Crystallographica Section F:Structural Biology Communications
• 2013, An Efficient Strategy for Small-Scale Screening and Production of Archaeal Membrane Transport Proteins in Escherichia coli, Plos One
• 2011, Crystal structure of archaeoglobus fulgidus CTP:Inositol-1-phosphate cytidylyltransferase, a key enzyme for di-myo-inositol-phosphate synthesis in (hyper)thermophiles, Journal of Bacteriology
• 2010, Production, crystallization and preliminary X-ray analysis of CTP:inositol-1-phosphate cytidylyltransferase from Archaeoglobus fulgidus, Acta Crystallographica Section F: Structural Biology and Crystallization Communications
• 2009, Structural and functional insights into sulfide:quinone oxidoreductase, Biochemistry
• 2006, Crystallisation and preliminary structure determination of a NADH: quinone oxidoreductase from the extremophile Acidianus ambivalens, Biochimica et Biophysica Acta - Proteins and Proteomics

Book chapter

• 2020, Structural biology techniques: X-ray crystallography, cryo-electron microscopy, and small-angle X-ray scattering
• 2019, Hydrogen Sulfide Signaling and Enzymology
• 2013, X-ray Crystallography

Conference paper

• Crosstalk between gasotransmitters at the H 2 S-synthesizing human cystathionine ß-synthase: heme-based regulation by CO and NO
• 2018-05, Structural insights into the regulation and inhibition of human cystathionine ß-synthase
• 2016-08, Quinone interaction on Type II NADH:quinone oxidoreductase from Staphylococcus aureus
• 2016-08, Investigating structural/function relationship in type II NADH:quinone oxidoreductases
• 2005-08-23, Structure of NADH: quinone oxidoreductase fromAcidianus ambivalens: electron entry point of aerobic respiratory chain

Conference poster

• 2023-10, AftD as a new potential drug target to fight tuberculosis, 13th ITQB NOVA PhD Students Meeting
• 2022-07-16, Crystallographic studies on enzymes involved in sulfide removal in the purple sulfur bacterium Thiocapsa roseopersicina, Second National Crystallographic Meeting
• 2022-05-19, Structural and functional insights into hydrogen sulfide homeostasis in pathogenic bacteria, 6th World Congress on Hydrogen Sulfide in Biology & Medicine