Building 8, Laboratory 2.32
Department of Biomedical Sciences and Medicine.
University of Algarve
Phone: +351 289 244 483
Eduardo Melo (Principal Investigator)
Evandro Tavares (MSc)
Joana Silva (MSc Student)
Cátia Correia (MSc)
About the PBBL
The Protein Biochemistry and Biophysics Laboratory (PBBL) addresses protein stability, protein folding/unfolding pathways and the implications of these regarding protein aggregation in degenerative disorders and disease. The protein biochemistry and biophysics laboratory applies engineering principles to understand, improve and dissect protein function(s). This laboratory has constructed novel fusions between fluorescent proteins and the prion protein that are suitable for expression at the membranes of targeted cell lines. These tools allow the analysis protein aggregation by confocal microscopy via Foster Resonance Energy Transfer (FRET). Biomolecules involved in oxidative protein folding in the endoplasmic reticulum and their implication in neurological disorders are studied within this group, with particular focus on biophysical approaches.
Konno, K., E. P. Melo, C. Lopes, C. F. Kaminski, I. Mehmeti, S. Lenzen, D. Ron, E. Avezov (2015) “ERO1-independent production of H2O2 within the endoplasmic reticulum fuels Prdx4 mediated oxidative protein folding”, J. Cell Biol. 211: 253-259.
Estrela, N., H. G. Franquelim, C. Lopes, E. Tavares, J. A. Macedo, G. Christiansen, D. E. Otzen, E. P. Melo* (2015) “Sucrose prevents protein fibrillation through compaction of the tertiary structure but hardly affects the secondary structure”
Proteins 83: 2039-2051.
Avezov, E., T. Konno, A. Zyryanova, W. Chen, R. Laine, A. Crespillo-Casada, E. P. Melo, R. Ushioda, K. Nagata, C. F. Kaminski, H. P. Harding, D. Ron (2015) “Retarded PDI diffusion and a reductive shift in poise of the calcium depleted endoplasmic reticulum”
BMC Biology 13: 2.
Tsunoda, S., E. Avezov, A. Zyryanova, T. Konno, L. Mendes-Silva, E. P. Melo, H. P. Harding, D. Ron (2014) “Intact protein folding in the glutathione-depleted endoplasmic reticulum implicates alternative protein thiol reductants”
eLife 3: e03421.
Tavares, E., J. A. Macedo, P. M. R. Paulo, C. Tavares, C. Lopes, E. P. Melo* (2014) “Live-cell FRET imaging reveals clustering of the prion protein at the cell surface induced by infectious prions” Biochim. Biophys. Acta – Molecular Basis of Disease 11: 744-751.
Brissos, V., N. Gonçalves, E. P. Melo, L. O. Martins (2014) “Directed evolution leads to aggregation-resistant variants with improved stability” PLOS One 9: e87209.
Rosa, M., C. Lopes, E. P. Melo, S. K. Singh, V. Geraldes, M. A. Rodrigues (2013). Measuring and modelling hemoglobin aggregation below freezing temperature J. Phys. Chem. B 117 8939 8946
Avezov, E., B. C. S. Cross, G. S. K. Schierle, M. Winters, H. P. Harding, E.P. Melo, C. F. Kaminski, D. (2013) Ron Lifetime imaging of a fluorescent protein sensor tracks ER thiol redox revealing its surprising stability J. Cell Biol. 201 337 349
Anjos, L., A. S. Gomes, E.P. Melo, A. V. M. Canário, D. M. Power. (2013) Cartilage acidic protein 2 a hyperthermostable, high affinity calcium-binding protein Biochim. Biophys. Acta – Proteins and Proteomics 1834 642 650
Fernandes, A.T., C. Lopes, L. O. Martins and E.P. Melo* (2012) Unfolding pathway of CotA-laccase and the role of cooper on the prevention of refolding through aggregation of the unfolded state Biochem. Biophys. Res. Commun 422 442 446
Fernandes, A.T., M. M. Pereira, C. S. Silva, P. F. Lindley, I. Bento, E. P. Melo* and L. O. Martins* (2011) “The removal of a disulfide bridge of CotA-laccase changes the slower motion dynamics involved in copper binding but has no effect on the thermodynamic stability” J. Biol. Inorg. Chem. 16: 641-651.
Melo, E.P., N. Estrela, C. Lopes, A. C. Matias, E. Tavares and V. Ochoa-Mendes (2010) “Compacting proteins: Pros and Cons od osmolyte-induced folding” Curr. Prot. Pep. Science 11: 744-751.
Zito, E., E.P. Melo, Y.Yang, Å. Wahlander, T.A. Neubert and D. Ron (2010) “Oxidative protein folding by an endoplasmic reticulum localized peroxiredoxin. Molec. Cell 40: 787-797.
Madeira, C. Estrela N., Ferreira JAB., Andrade SM., Costa, SMB., Melo EP. (2009), “Fluorescence lifetime imaging microscopy and fluorescence resonance energy transfer from cyan to yellow fluorescent protein validates a novel method to cluster proteins on solid surfaces”, Journal of Biomed Optics, 14(4):044035.
Morgado, I., Fernandes, AT; Martins, LO; Melo, EP (2009) “The hyperthermophilic nature of the metallo-oxidase from Aquifex aeolicus”, Biochim. Biophys. Acta- Proteins and Proteiomics 1794(1): 75-83.
E.P. Melo, E. Lundberg, N.L. Estrela, A.E. Samer-Eriksson and D. Power (2008) “Piscine transthyretin hormone affinity and fibril formation: The role of the N-terminal”, Mol. Cell. Endocrinol. 295: 48-58.
Brissos, V., E.P. Melo, J.M.G. Martinho and J.M.S. Cabral (2008) “Biochemical and Structural Characterisation of Cutinase Mutants in the Presence of the Anionic Surfactant AOT”, Biochim. Biophys. Acta 1784: 1326-1334.
Baptista, R.P., S.H. Pedersen, G.J.M. Cabrita, D.E. Otzen, J.M.S. Cabral and E.P. Melo (2008) “Thermodynamics and Mechanism of Cutinase Stabilization by Trehalose” Biopolymers 89(6):538-547.
Durão, P., Z. Chen, A.T. Fernandes, M.M. Pereira, E.P. Melo, and L.O. Martins (2008) “Copper Incorporation Into Recombinant CotA-laccase from Bacillus subtilis: Characterization of Cu Full-Loaded Enzymes”. J. Biol. Inorg. Chem. 13 (2):183-193.
Stable clones expressing the fusions PrP-CFP-GPI (A), PrP-YFP-GPI (B) and both PrP-CFP-GPI and PrP-YFP-GPI (double clone) (C). Colocalization for the fusions PrP-CFP-GPI and PrP-YFP-GPI is shown in the plot in (C) with a Pearson’s coefficient of 0.75. (D) Cell substructure of the double clone showing the nuclei labeled with Hoechst 33342 in blue and the ER labeled with ER-tracker red in red (left panel). The image on the right shows a tridimensional view of the double clone, obtained after deconvolution and SFP rendering with the Huygens software (SVI, The Netherlands), to highlight the non-homogeneous distribution of fluorescent proteins on the membrane where clusters of fluorescent proteins are visible in green. The ER is shown in red. (E) Neuroblastoma cells (left panel PK1 and right panel PK1 knockdown cells) stained with mouse anti-PrP monoclonal antibody POM1 and Alexa Fluor 647-labeled goat anti-mouse IgG1 for endogenous PrP (red) and counterstained with Hoechst 33342 for nuclei (blue). Scale bars correspond to 5 μm.
PTDC/CTM-NAN/2700/2012 – “Nano-Dímeros Metálicos para Biosensores de Proteína Individual com Resposta Plasmónica Intensificada”, Team member
PTDC/QUI-BIQ/119677/2010 – “A biophysical approach to oxidative protein folding in the endoplasmic reticulum”, Principal Investigator
PTDC/EQU-EQU/104318/2008 – “Controlled freezing to improve the stability and delivery of therapeutic proteins”, Team member.
PTDC/QUI/73027/2006 – “Single-Molecule Detection of the Prion Protein: Characterization of Conformational Changes Involved in Prion Diseases”, Coordinator.
POCI/CVT/56668/2004 “A Murine Neuroblastoma Cell-Based Approach to Study and Prevent Prion Diseases”, Principal Investigator.
PTDC/QUI/64658/2006 – “Single Molecule Signalling of peptide metal binding”. Team member
PTDC/QUI/64112/2006 – “Transferência Electrónica fotoinduzida por proteínas hémicas artificiais em sistemas de nanotubos de carbono”. Team member.
POCI/QUI/58816/2004 “Lipid-Lipid and Protein-Lipid Interactions in Model Systems of Cell Membranes”. Team member.
POCI/QUI/57387/2004 “Molecular Organization of Functionalized Porphyrins in Giant Vesicles and Supramolecules” . Team member.
Sapiens 2002, Portugal. “Relation Between CYP3A4 Gene Polymorphism, Protein Structure and Activity in Disease Setting”, Team Member.
Sapiens 2002, Portugal. “Design of Stabilizing Affinity Molecular Probes: A Practical Approach to Assess and Enhance Protein Stability”, Team Member.