Cell Wall Biogenesis in bacteria
The cell wall is one of the hallmarks of bacterial cells. It is a very robust structure that helps bacteria to maintain their shape, regulates their division and allows them to invade and colonize our body. Many of our most successful antibiotics target proteins that maintain the bacterial cell wall, thus causing its disruption and eventually cell death.
I want to discover the mechanisms proteins use to construct the bacterial cell wall for two reasons; understand how these remarkable creatures operate; and figure out new ways to combat the ones that infect us causing morbidity and mortality.
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Responsibilities:
• Design of experiments (DoE) and multivariate analysis.
• Protein engineering.
• Supervise undergraduate and graduate students.
• Manage the experimental progress of the project.
• Set up novel assays and bring new ideas for the development of the project.
• Communicate results in internal meetings and at international conferences.
• Attract competitive funding.
Drug discovery for trypanosomatid parasites.
Early target-based drug discovery for the major human and animal trypanosomatid parasites. These cause a variety of diseases such as Sleeping Sickness (Trypanosome brucei rhodesiense/gambiense), Chagas Disease (T. cruzi), Visceral Leishmaniasis (Leishmania donovani) and Nagana (T. vivax, T. congolense) in domestic animals. The common target for drug intervention in all these parasites is RNA Editing Ligase 1 (REL1), an essential protein for the maturation of mitochondrial mRNA and survival of the parasites. Studying this protein will prove to be pivotal for finding ways to target all these parasites. However, its biochemical and biophysical characterisation is extremely difficult as it is an exceptionally unstable protein. My effort focuses on engineering derivatives that are going to be stable, functional and amenable to biochemical and biophysical characterisation.
Responsibilities:
• Design of experiments (DoE) and multivariate analysis.
• Protein engineering to stabilise extremely unstable protein molecules.
• Trying to obtain high resolution 3D crystal structures of RNA Editing Ligase 1 in complex with hit/lead compounds.
• Setting up pipelines for the validation of of hit compounds generated from HTS campaigns, using ortholog protein targets, biochemical and biophysical technologies.
• Supervise undergraduate and graduate students.
• Manage the experimental progress of the project.
• Set up novel assays and bring new ideas for the development of the project.
• Communicate results in internal meetings and at international conferences.
• Attract competitive funding.
Protein secretion through the Sec system.
Dissecting the mechanisms bacteria use to sort and target proteins destined to be secreted to the cytoplasmic membrane and beyond. My work led to discovering that the mature domains of preproteins contain targeting signals that direct them to the Sec secretory pathway. We discovered that the initial docking site on the membrane receptor (SecA-SecYEG complex) consists of hydrophobic patches. Furthermore, the flexibility of the preprotein is the factor that determines the efficiency of its secretion through the Sec system.
Responsibilities:
• Design of experiments (DoE) and multivariate analysis.
• Engineer proteins to test various working hypotheses about the mechanism of preprotein secretion.
• Supervise undergraduate and graduate students.
• Manage the experimental progress of the project.
• Set up novel assays and bring new ideas for the development of the project.
• Communicate results in internal meetings and at international conferences.
Modulation of immune response in mammals.
Soluble MHC antigens are detected in body fluids but their role and origin are still unclear. This study examined whether serum IA antigens, isolated from BALB/c mice (sIA(d)), could modulate the immune response. Specific purification procedures isolated intact IA molecules, which were thereafter applied to functional assays. Our results suggest that sIA(d), following steps similar to surface class II antigen binding mechanisms, stimulate the initiation of a humoral or cellular immune response but rather inhibit the effector phase of the reactions, attributing thus soluble class II MHC antigens important immunomodulatory roles. The results of this study were published in the peered review journal Immunobiology (Sardis et al., 2010 Mar;215(3):194-205).
Responsibilities:
• Design of experiments (DoE) and multivariate analysis.
• Set up novel assays and bring new ideas for the development of the project.
• Manage the experimental progress of the project.
• Communicate results in internal meetings and at international conferences.
• B.Sc. research thesis preparation.