25–30 May 2020, Spetses Island, Greece
Presently, I am a cell and molecular biology lecturer at the School of Health Sciences and Technologies, Lusófona University, and I am a member of the CBIOS group. At CBIOS I am focused on studying the impact of pharmacological ROS modulation (mainly via SOD mimics) on cell migration and adhesion in the context of cancer and metastasis. This work is done in collaboration with several national and international collaborators. In addition, I am also continuing my previous research projects on the impact of GAAP-induced ROS accumulation in cell invasion with Prof Geoffrey Smith at Cambridge University and Prof Maddy Parsons at Kings College.
My current research interests involve exploring the impact of Ca2+ and ROS in the cellular mechanisms underlying cell migration, adhesion and invasion. Both Ca2+ and ROS levels are highly regulated and can be affected by many cellular events and signalling pathways. I am particularly interested in exploring how these processes affect cell migration and adhesion pathways, the dynamics of the cell’s cytoskeleton and adhesion structures and how this in turn affects the cell ability to migrate and invade. This area is of great importance to the integrated comprehension of the molecular mechanisms underlying the mechano-biological properties of the cell and the dynamics of the cellular structures involved.
Throughout my research career I have worked with and developed several complex molecular reporters and cellular imaging techniques to explore cell migration and structure, FA dynamics, protein activity localization, Ca2+ and ROS measurements protein interaction, etc.
These technical skills include:
1. Protein, RNA and DNA handling
• PCR, RT-qPCR, sequence analysis
• Protein analysis: SDS-PAGE, Western blot, immunofluorescence, protein-protein interaction (complementation assays, IP, non denaturing PAGE, FRET), molecular exclusion chromatography.
• Membrane and soluble protein purification
2. Genetic engineering
• Cloning, sub-cloning, fusion protein, directed mutagenesis, subcellular targeting of recombinant proteins.
• Generation of constitutive or inducible gene KD or gene overexpressing mammalian cell lines.
• Genetically encoded biosensors for live cell imaging: protein-protein interaction, [Ca2+], [H2O2] and protease activity.
3. Cell (eukaryotic and prokaryotic) culture, handling and analysis
• Mammalian cell lines, yeast and bacteria manipulation, culture
• Transduction and transfection methods.
• Apoptosis. Cell migration and invasion. Gene reporter assays. Intracellular Ca2+ measurements.
• Microscopy: wide field, confocal, live cell time lapse, FRET, FRAP, TIRF, IRM.
• Flow cytometry analysis and cell sorting.
4. Virology techniques
• 7 years of experience in protocol and usage of BSL 2 and 3 (lentiviral vectors, poxvirus and HIV).
• Virus infection, production, titration, selection, concentration and purification.
Geneticaly encoded biosensors
Saraiva N, Prole D, Carrara G, Johnson B, Taylor W, Parsons M and Smith G. hGAAP promotes cell adhesion and migration via the stimulation of store-operated Ca2+ entry and calpain 2. J Cell Biol 2013. Aug 19;202(4):699-713.
Fernandes AS, Flórido A, Saraiva N, Cerqueira S, Ramalhete S, Cipriano M, Cabral M, Miranda J, Castro J, Costa J, Oliveira N. Role of the copper(II) complex CupyN5 in intracellular ROS and breast cancer cell motility and invasion. Chem Biol Drug Des. 2015 Oct;86(4):578-88.
Carrara G, Saraiva N, Parsons M, Byrne B, L. Prole D, Taylor C, and Smith G. Golgi anti-apoptotic proteins are highly conserved ion channels that affect apoptosis and cell migration. J Biol Chem. 2015 May 1;290(18):11785-801.
Saraiva N, Prole D, Carrara G, Maluquer de Motes C, Johnson B, Byrne B, Taylor W and Smith G. Human and viral Golgi anti-apoptotic proteins (GAAPs) oligomerize via different mechanisms and monomeric GAAP inhibits apoptosis and modulates calcium. J Biol Chem. 2013 May 3;288(18):13057-67.
Carrara G, Saraiva N, Gubser C, Johnson B and Smith G. A six transmembrane topology for Golgi Anti-Apoptotic Protein (GAAP) and Bax Inhibitor 1 (BI-1) provides a model for the Transmembrane Bax Inhibitor Containing Motif (TMBIM) family. J Biol Chem. 2012 May 4;287(19):15896-905.
25–30 May 2020, Spetses Island, Greece