|
Ph.D. in Pharmaceutical and Clinical Biochemistry (FCUL, Portugal)
Lic. in Biochemistry (FCUL, Portugal)
My research activity has been focused on:
• Reactive oxygen and nitrogen species (ROS/RNS) metabolism
– Mammalian formation/consumption of ROS/RNS,
nitric oxide radical, superoxide anion radical and peroxides (hydrogen and lipidics)
– Mechanisms of ROS/RNS-mediated diseases,
namely in the pathologies associated with vascular dysfunction and alcohol hepatotoxicity
• Human signalling pathways
– Novel pathways of formation/consumption of signalling molecules, namely nitric oxide
– In vitro studies with purified rat liver and human liver xanthine oxidase, xanthine
dehydrogenase and aldehyde oxidase
– In situ studies with human cell lines
• New catalytic activities of molybdoenzymes
– NADH oxidation, nitrite reduction, nitric oxide formation, carbon dioxide reduction (carbon dioxide scavenging)
– Mechanistic and kinetic studies
– Mammalian and bacterial enzymes
• Structure-activity relationships in metalloenzymes
– Mechanistic and kinetic studies
– Small proteins mimetics (molybdenum-substituted rubredoxin)
– Mammalian and bacterial enzymes (rat and human liver xanthine oxidase, xanthine
dehydrogenase and aldehyde oxidase, bovine xanthine oxidase, Desulfovibrio aldehyde
oxidoreductases and formate dehydrogenases, Marinobacter nitric oxide reductase)
Methodologies used:
• Optimization of enzyme purification procedures
– Chromatography (ionic exchange, adsorption, filtration), centrifugation, precipitation, electrophoresis
• Kinetic characterisation
– Reduction/oxidation of nitric oxide precursors, nitric oxide, metabolites relevant to
ischemia injury, alcohol hepatotoxicity (purinic and pyridinic compounds), aldehydes
metabolism (aromatic and aliphatic), carbon dioxide metabolism
– Spectroscopic (UV-visible, fluorescence, EPR, NMR), HPLC and polarographic methodologies
• Mechanistic studies
– Enzymes redox active centres characterisation, electron transfer within the centres and
substrates/products/inhibitors, aiming to characterise the reactions mechanism with atomic detail
– EPR, NMR, mass, UV-visible spectroscopies,
with substrates, products, specific inhibitors, isotopically labelled compounds and spin-traps
• Quantification of enzymatic ROS/RNS formation (•NO, O2•-, H2O2, HO•, ONOO-),
endogenous antioxidants (vitamins E and C, GSH, SOD, catalase),
and oxidative modifications on proteins, nucleic acids and lipids
– Spectrophotometric, fluorimetric, EPR, MS, HPLC, GC and electrophoretic methodologies,
lipid purification techniques
• Tissue cultures (human cell lines)
– Fluorescence methodologies (probes to follow signalling molecules)
• Models of oxidative stress
|
|
