Universidade Nova de Lisboa - REQUIMTE - FCT

Main Research Topics

BIOIN / BIOPROT (BioINorganic and Biophysical PROtein Groups)
(Sofia R. Pauleta, Isabel Moura, José J.G. Moura)

- Paramagnetic NMR – Metalloproteins – Transition Metals

Application of NMR techniques to study metalloproteins containing transition metals at active sites, focusing on paramagnetic effects. Case studies: electron transfer proteins (cupredoxins, heme and iron-sulfur proteins) and metalloenzymes. Correlations with EPR and Mössbauer spectroscopies. (51)V NMR to study interactions between G-actin and decavanadate.

R. M. Almeida, S. R. Pauleta, I. Moura, J.J.G. Moura. Rubredoxin as a paramagnetic relaxation-inducing probe. 2009. J. Inorg. Biochem. 103: 1245-53.

S. Ramos, M. Manuel, T. Tiago, R. Duarte, J. Martins, C. Gutiérrez-Merino, Jose J.G. Moura, M. Aureliano. Decavanadate interactions with actin: inhibition of G-actin polymerization and stabilization of decameric vanadate. 2006. J. Inorg. Biochem. 100: 1734-43.

- Protein NMR, Protein Solution 3D Structure
Application of NMR techniques to the determination of solution structure of metalloproteins and transcription regulators. Structural and mechanistic studies of metalloenzymes, electron transfer complexes and transcription regulators using different biochemical and biophysical techniques (e.g., NMR spectroscopy, microcalorimetry).

S. R. Pauleta, A.G. Duarte, M.S. Carepo, A.S. Pereira, P. Tavares, I. Moura, J.J.G. Moura. NMR assignment of the apo-form of a Desulfovibrio gigas protein containing a novel Mo-Cu cluster. 2007. Biomolecular NMR Assignments. 1 (1), 81-3.

- Protein-Protein Interactions
Structure of electron transfer complexes using NMR data and molecular docking algorithm (BiGGER) and other structural constraints (e.g., site directed mutagenesis, green pathways, electron transfer kinetics).
S. Dell’Acqua, S. R. Pauleta, E. Monzani, A.S. Pereira, L. Casella, J.J.G. Moura, I. Moura. Pseudomonas nautica cytochrome c552 is the electron donor to nitrous oxide reductase (N2OR). 2008. Biochemistry. 47 (41), 10852-62.

- Medical Applications
NMR detection of biological markers in oncological conditions (under the protocol with IPO, Dra. M. Roldão).

(Bio)molecular Structure and Interactions
(Eurico J. Cabrita)

In a broad context the research is focused in the application and development of NMR techniques to study intermolecular interactions in chemical and biological systems and to the conjugation of NMR, molecular modeling and organic synthesis as a tool to the study of chemical and biochemical reaction mechanisms.

- Study of intermolecular interactions in non-conventional solvents (ionic Liquids and scCO2 - High pressure NMR)
Application of NMR techniques, with special focus to HOESY and diffusion NMR, to the study of intermolecular interactions in two of the most promising alternative media to volatile organic compounds (VOCs), namely, ionic liquids (ILs) and supercritical (sc)CO2, in order to establish a molecular based interpretation of their solvent properties;

G. I. Ivanova, E. R. Vão, M. Temtem, A. Aguiar-Ricardo, T. Casimiro, E. J. Cabrita, High-pressure NMR characterization of triacetyl-beta-cyclodextrin in supercritical carbon dioxide, Magn. Reson. Chem., 2009, 47, 133-141, DOI: 10.1002/mrc.2365

M. Temtem, T. Casimiro, A. G. Santos, A. L. Macedo, E. J. Cabrita*, A. Aguiar-Ricardo, Molecular interactions and CO2-philicity in supercritical CO2. A high-pressure NMR and molecular modeling study of a perfluorinated polymer in scCO2, J. Phys. Chem. B, 2007, Vol. 111, Nr. 6, 1318-1326. DOI: 10.1021/jp0660233

- Study of intermolecular interactions in chemical systems: catalysis and asymmetric chemistry
Application of NMR techniques to the rationalization of reaction mechanisms with special focus to asymmetric chemistry and catalysis;

S. M. Bakalova, F. J. S. Duarte, M. K. Georgieva, E. J. Cabrita, A. G. Santos, An alternative mechanism for Diels-Alder reactions of Evans auxiliary derivatives, Chem. Eur. J. 2009, , http://dx.doi.org/10.1002/anie.200900628

- Study of intermolecular interactions in biological systems
Application of NMR techniques (DOSY, STD-NMR, trNOE) to the study of protein-ligand interactions in a biocatalysis or "drug-design" context;

A. Viegas, N. F. Brás, N. M. F. S. A. Cerqueira, P. A. Fernandes, J. A. M. Prates, C. M. G. A. Fontes, M. Bruix, M. J. Romão, A. L. Carvalho, M. J. Ramos, A. L. Macedo, E. J. Cabrita, Molecular determinants of ligand specificity in family 11 Carbohydrate Binding Modules (CBM11): a NMR, X-Ray crystallography and computational chemistry approach, FEBS Journal, 2008, 275, 2524-2535. doi:10.1111/j.1742-4658.2008.06401.x

- Structure elucidation and complex mixture analysis
G. Ivanova, L. S. Serafim, P. C. Lemos, A. M. Ramos, M. A. M. Reis, E. J. Cabrita, Influence of feeding strategies of mixed microbial cultures on the chemical composition and microstructure of copolyesters P(3HB-co-3HV) analyzed by NMR and statistical analysis, Magn. Reson. Chem., 2009, 47, 497-504, DOI 10.1002/mrc.2423

G. I. Ivanova, E. J. Cabrita, R. O’Connor, A. J. Eustace, D. F. Brougham, Application of diffusion-ordered spectroscopy for the analysis of cancer related biological samples, Bulg. Chem. Comm., 2008, 40, 464-468.

Heme Proteins NMR
(Carlos Salgueiro)

Application of NMR techniques to the study of Biological Systems, and particularly those that involve electron transfer metalloproteins (multihaem cytochromes, ferredoxins, etc).
Structural-functional relationships in electron transfer and elucidation of electron transfer mechanisms involved in metal reduction by biological molecules.
The main area of experimental work includes, among others:

- Protein solution structure determination by NMR,

Morgado, L., Bruix, M. Londer, Y.Y., Pokkuluri, P.R., Schiffer, M. & Salgueiro, C.A. (2007) Redox-linked conformational changes of a multiheme cytochrome from Geobacter sulfurreducens. Biochem. Biophys. Res. Commun. 360, 194–198.
Paixão, V.B., Salgueiro, C.A., Brennan, L., Reid, G.A., Chapman, S_K., Turner, D.L. (2008) Solution structure of a tetrahaem cytochrome from Shewanella frigidimarina reveals a novel family structural motif, Biochemistry 47,11973-11980.

- folding and unfolding protein studies by NMR,

Todorovic, S., Leal, S.S., Salgueiro, C.A., Zebger, I., Hildebrandt, P., Murgida, D.H. & Gomes, C.M. (2007) A spectroscopic study of the temperature induced modifications on ferredoxin folding and iron-sulfur moieties, Biochemistry 46,10733-10738.

Structural Biology
(Anjos L. Macedo)

The main area of experimental work includes, among others:

- Protein structural studies and ligand protein interactions in heme binding proteins,

J.S. Dias, A.L.Macedo, G. C. Ferreira, F. C. Peterson, B.F. Volkman, B. J. Goodfellow (2006),The first Structure of a protein from the Soul/HBP family: Murine p22HBP, J. Biol Chem., 281, 31553-61.

- rationalization of molecular interactions in ribotoxins and unfolding protein studies by NMR,

Viegas A, Herrero-Galán E, Oñaderra M, Macedo AL and Bruix M, (2009)Solution structure of hirsutellin A – new insights into the active site andinteracting interfaces of ribotoxins, FEBS Journal 276, 2381–239.