Universidade de Aveiro - DQ

Departamento de Química Universidade de Aveiro Campus Universitário de Santiago 3810-193 Aveiro Portugal

Tel: +351 234 370 360 Fax: +351 234 370 084

Main Research Topics

João Rocha - Solid-State NMR [refs 1-13]

From 1988 to 1995 we have pioneered the development and application of certain techniques for the study of quadrupolar nuclei, such as 2H exchange spectroscopy, quadrupole nutation and double rotation, and satellite transition spectroscopy [1-3]. From 1995 on we focused on half-integer quadrupolar nuclei and had a lot of fun with multiple-quantum (MQ) MAS NMR [4] and related techniques, such as FAM MQ MAS [5], HETCOR MQ MAS [6] and ST MAS, I-ST MAS [7], including SPAM-multiplex methods [8], and 3D 27Al-31P MQ-HMQC [9]. Much of this work was concentrated on inorganic materials such as clays and layered solids, zeolites and other micro and mesoporous materials. In the last few years we became interested in using high-resolution 1H CRAMPS techniques (FSLG, PMLG, DUMBO) for studying organic-inorganic hybrid materials [10-13]. We have just started work on small proteins. In 2007 we initiated in Aveiro (with F. Taulelle) the international event ‘SMARTER - Structure elucidation by coMbining mAgnetic Resonance, compuTation modEling and diffRactions’, bridging scientific communities. SMARTER 2 was held in Aveiro in 2011 and the third was held in Paris 2012.

Luís Mafra - Solid-State NMR [refs 13-22]

Methodological development and applications in solid-state NMR using high-resolution methods for 1H (using Combining Rotation And Multiple Pulse Sequences (CRAMPS) techniques) and quadrupolar spins to study small molecules and inorganic-organic hybrid materials [13-22]. 1D and 2D heteronuclear and homonuclear high-resolution 1H Lee-Goldburg based CRAMPS NMR techniques in solids (e.g., FSLG, LG-CP) have been first implemented in Portugal by him since 2004 (during his PhD) [14]. In particular, modern 1H homonuclear decoupling schemes such as PMLG and DUMBO/eDUMBO have been exploited to study various systems of biological interest [14-18]. During the last years we have explored the fast MAS technology (up to 67 kHz) in combination with CRAMPS to enhance the resolution of 1H NMR spectra [16], including the improvement of 1H-1H Double-Quantum (DQ) experiments using RN symmetry-based recoupling pulse sequences [17] and 1H-quadrupolar spins correlation methods [13]. His latest research lines of interest resides in i) the study of intermolecular interactions in porous materials with interesting gas sorption (e.g., CO2) and catalytic properties [19] and, very recently, in ii) structural elucidation of biosolids (e.g., biomacromolecules) and iii) combining high-resolution solid-state NMR at high magnetic fields, X-ray diffraction and computational approaches to study packing interactions (e.g., weak CH∙∙∙X and CH∙∙∙p / strong hydrogen bonds and p∙∙∙p interactions) on pharmaceutical hydrates/anhydrates systems [18]. He has recently contributed with a chapter in the Encyclopedia of Nuclear Magnetic Resonance related with molecular sieves [22].

Artur Silva - Liquid-State NMR: Small Molecules [refs 23-33]

NMR is being used as the main tool for the comprehensive characterization of the chemical structure, conformations and configurations of synthetic and natural molecules. My interests are: a) Synthesis and structural characterization of oxygen (mainly polyphenolics) and nitrogen heterocyclic compounds; b) Structural characterization of tetrapyrrolic macrocycles; c) Chemical composition of plants; d) Structural characterization of polyphenolic compounds isolated from red wines.

Ana Gil - Liquid state NMR, HRMAS NMR and hyphenated NMR (LC-NMR and LC-NMR/MS); Solid-State NMR [refs 34-50]

One of my main present interests is the metabonomic analysis human biofluids and biological tissues for the biochemical characterization, early diagnosis, prognosis and monitoring of diseases and treatment, mainly focused on several prenatal diseases (analysis of amniotic fluid, maternal blood and urine, newborn urine, umbilical cord blood, placenta tissue) and in lung cancer (analysis of blood, urine and biopsy tissues) [34-39]. This work involves state of the art analytical strategies based on high resolution NMR (liquids, HRMAS, hyphenated NMR, LipoProfiling for lipoprotein analysis) and MS technologies (namely UPLC-MS and targeted LC-MS for lipoprofiling). A similar strategy is being applied in our group to the understanding of the metabolic effects of drugs (both established and novel) and drug delivery agents (e.g. polymers and biopolymers, nanoparticles) on cellular metabolism [40, 41]. Models being studied comprise human osteosarcoma and hepatocyte cell lines and external stimuli include anti-tumoral drugs and specific polysaccharides used as drug delivery vehicle [42-44]. An additional line is also emerging on the effects of environmental effects (e.g.metal contaminants, pesticides) on cellular metabolism.
A parallel interest relates to food analysis, again employing a metabonomics strategy, here put to use for the routine quality control of products and processes in the food industry (brewing industry, beverages and wine industries) [45-47].

Concomitant interests reside in solid samples structural and dynamics study, mainly through solid state NMR and namely focusing on structure/dynamics characterization of a) solid mono- and oligosaccharides and inclusion complexes; characterization of hydrogen-bonding networks; and b) biopolymers e.g. plant suberin [48, 49] and dextran-based polysaccharide systems [50].

Brian Goodfellow - Liquid-State NMR: Heme Metabolism; Bioinorganic Chemistry; Metabolomics [refs 36-39, 51-55]

I am interested in proteins involved in heme metabolism and have studied the presequence of the first protein involved in heme biosynthesis, 5-amino levulinic acid synthase [51], and have recently determined the solution structure and heme-binding site of murine p22HBP [52]. Here we have identified a novel fold and binding site not seen before in a eukaryotic protein. The function of this protein is unknown. I am also interested a related protein SOUL which has been found to be a novel BH3 domain containing protein [53].

I am also involved in the study of small metalloproteins containing simple Fe-S centres to probe how these centres are stabilized by, in particular, hydrogen bonds. By replacing the native Fe by diamagnetic metals, such as Zn and Cd the structure of these proteins can be obtained and by using paramagnetic derivatives, such as Ni, the electronic distribution at the active site can be probed using residual dipolar coupling and pseudocontact shifts [54, 55].

The use of NMR based metabolomics is also an area of interest, especially as applied to biofluids for disease diagnosis and prediction in the area of prenatal health [36-39].

Iola Duarte - Liquid-State NMR and HRMAS; Metabonomics [refs 37, 38, 44, 56, 57]

A main research topic consists of the application of High Resolution NMR methods, in tandem with multivariate statistics (chemometrics), to the analysis of complex biological samples, namely biofluids, tissues and cells, aiming at the discovery of metabolic fingerprints of a certain physiological status, disease or response to external stimuli (e.g. drug). Specific research lines comprise:
- Investigation of altered cancer metabolism and development of new diagnostic methods based on NMR metabonomics of tissues and biofluids (e.g. blood plasma and urine).
- Characterization of the metabolic responses of human cultured cells to conventional and novel anticancer agents.
- Development of cell metabonomics and isotopomer-assisted metabolic profiling strategies for assessing the toxicity mechanisms and sub-toxic biological effects of biomaterials, namely nanomedicines developed for drug delivery and other biomedical applications.
- Characterization of the metabolic responses of vegetal and animal models to stress/toxic insults.

Selected Publications

1. Duer, MJ and Rocha, J, "A two-dimensional solid-state 2H exchange NMR study of the molecular motion in the kaolinite:DMSO intercalation compound". J. Magn. Reson., 98: 524 (1992).

2. Rocha, J, Klinowski, J, Barrie, PJ, Jelinek, R, and Pines, A, "Solid-state 27Al NMR studies of aluminophosphate molecular sieves: enhanced resolution by quadrupole nutation and double-rotation". Solid State Nucl. Magn., 1: 217 (1992).

3. Jager, C, Rocha, J, and Klinowski, J, "High-speed satellite transition 27Al MAS NMR spectroscopy". Chem. Phys. Lett., 188: 208 (1992).

4. Anderson, MW, Agger, JR, Luigi, DP, Baggaley, AK, and Rocha, J, "Cation sites in ETS-10: 23Na 3Q MAS NMR and lattice energy minimisation calculations". Phys. Chem. Chem. Phys., 1: 2287 (1999).

5. Morais, CM, Lopes, M, Fernandez, C, and Rocha, J, "Assessing the potential of fast amplitude modulation pulses for improving triple-quantum magic angle spinning NMR spectra of half-integer quadrupolar nuclei". Magn. Reson. Chem., 41: 679 (2003).

6. Fernandez, C, Morais, C, Rocha, J, and Pruski, M, "High-resolution heteronuclear correlation spectra between 31P and 27Al in microporous aluminophosphates". Solid State Nucl. Magn., 21: 61 (2002).

7. Amoureux, JP, Morais, C, Trebosc, J, Rocha, J, and Fernandez, C, "I-STMAS, a new high-resolution solid-state NMR method for half-integer quadrupolar nuclei". Solid State Nucl. Magn., 23: 213 (2003).

8. Malicki, N, Mafra, L, Quoineaud, AA, Rocha, J, Thibault-Starzyk, F, and Fernandez, C, "Multiplex MQMAS NMR of quadrupolar nuclei". Solid State Nucl. Magn., 28: 13 (2005).

9. Morais, CM, Montouillout, V, Deschamps, M, Iuga, D, Fayon, F, Paz, FAA, Rocha, J, Fernandez, C, and Massiot, D, "1D to 3D NMR study of microporous alumino-phosphate AlPO4-40". Magn. Reson. Chem., 47: 942 (2009).

10. Mafra, L, Rocha, J, Fernandez, C, and Paz, FAA, "Characterization of microporous aluminophosphate IST-1 using 1H Lee-Goldburg techniques". J. Magn. Reson., 180: 236 (2006).

11. Cunha-Silva, L, Mafra, L, Ananias, D, Carlos, LD, Rocha, J, and Paz, FAA, "Photoluminescent lanthanide-organic 2D networks: A combined synchrotron powder X-ray diffraction and solid-state NMR study". Chem. Mater., 19: 3527 (2007).

12. Mafra, L, Gomes, JRB, Trebosc, J, Rocha, J, and Amoureux, JP, "1H-1H double-quantum CRAMPS NMR at very-fast MAS nR = 35 kHz): A resolution enhancement method to probe 1H-1H proximities in solids". J. Magn. Reson., 196: 88 (2009).

13. Siegel, R, Rocha, J, and Mafra, L, "Combining STMAS and CRAMPS NMR spectroscopy: High-resolution HETCOR NMR spectra of quadrupolar and 1H nuclei in solids". Chem. Phys. Lett., 470: 337 (2009).

14. Coelho, C, Rocha, J, Madhu, PK, and Mafra, L, "Practical aspects of Lee-Goldburg based CRAMPS techniques for high-resolution 1H NMR spectroscopy in solids: Implementation and applications". J. Magn. Reson., 194: 264 (2008).

15. Mafra, L, Coelho, C, Siegel, R, and Rocha, J, "Assessing the performance of windowed 1H CRAMPS methods, on biological solids, at high-field and MAS up to 35 kHz". J. Magn. Reson., 197: 20 (2009).

16. Siegel, R, Mafra, L, and Rocha, J, "Improving the 1H indirect dimension resolution of 2D CRAMPS NMR spectra: A simulation and experimental investigation". Solid State Nucl. Magn., 39: 81 (2011).

17. Mafra, L, Siegel, R, Fernandez, C, Schneider, D, Aussenac, F, and Rocha, J, "High-resolution 1H homonuclear dipolar recoupling NMR spectra of biological solids at MAS rates up to 67 kHz". J. Magn. Reson., 199: 111 (2009).

18. Mafra, L, Santos, SM, Siegel, R, Alves, I, Paz, F, Dudenko, D, and Spiess, HW, "Packing interactions in hydrated and anhydrous forms of the antibiotic Ciprofloxacin: a solid-state NMR, X-ray diffraction and computer simulation study". J. Amer. Chem. Soc.: (in press).

19. Pinto, ML, Mafra, L, Guil, JM, Pires, J, and Rocha, J, "Adsorption and Activation of CO2 by Amine-Modified Nanoporous Materials Studied by Solid-State NMR and 13CO2 Adsorption". Chem. Mater., 23: 1387 (2011).

20. Mafra, L, Rocha, J, Fernandez, C, Castro, GR, Garcia-Granda, S, Espina, A, Khainakov, SA, and Garcia, JR, "Characterization of layered gamma-titanium phosphate (C2H5NH3)[Ti(H1.5PO4)(PO4)]2.H2O intercalate: A combined NMR, synchrotron XRD, and DFT calculations study". Chem. Mater., 20: 3944 (2008).

21. Garcia-Granda, S, Khainakov, SA, Espina, A, Garcia, JR, Castro, GR, Rocha, J, and Mafra, L, "Revisiting the Thermal Decomposition of Layered gamma-Titanium Phosphate and Structural Elucidation of Its Intermediate Phases". Inorg. Chem., 49: 2630 (2010).

22. Klinowski, J and Mafra, L, Crystalline molecular sieves, in Encyclopedia of Nuclear Magnetic Resonance, R.K. Harris and R. Wasylishen, Editors. in press, Wiley.

23. Oliva, CG, Silva, AMS, Resende, D, Paz, FAA, and Cavaleiro, JAS, "Highly Enantioselective 1,4-Michael Additions of Nucleophiles to Unsaturated Aryl Ketones with Organocatalysis by Bifunctional Cinchona Alkaloids". Eur. J. Org. Chem.: 3449 (2010).

24. Esteves, CIC, Santos, CMM, Brito, CM, Silva, AMS, and Cavaleiro, JAS, "Synthesis of Novel 1-Aryl-9H-xanthen-9-ones". Synlett: 1403 (2011).

25. Oliveira, J, Mateus, N, Rodriguez-borges, JE, Cabrita, EJ, Silva, AMS, and de Freitas, V, "Synthesis of a new pyranoanthocyanin dimer linked through a methyl-methine bridge". Tetrahedron Lett., 52: 2957 (2011).

26. Carvalho, CMB, Neves, M, Tome, AC, Paz, FAA, Silva, AMS, and Cavaleiro, JAS, "1,3-Dioxopyrrolo[3,4-b]porphyrins: Synthesis and Chemistry". Org. Lett., 13: 130 (2011).

27. He, JR, Oliveira, J, Silva, AMS, Mateus, N, and De Freitas, V, "Oxovitisins: A New Class of Neutral Pyranone-anthocyanin Derivatives in Red Wines". J. Agric. Food Chem., 58: 8814 (2010).

28. Kijjoa, A, Santos, S, Dethoup, T, Manoch, L, Almeida, AP, Vasconcelos, MH, Silva, A, Gales, L, and Herz, W, "Sartoryglabrins, Analogs of Ardeemins, from Neosartorya glabra". Nat. Prod. Commun., 6: 807 (2011).

29. Neri, DFM, Balcao, VM, Cardoso, SM, Silva, AMS, Domingues, MDM, Torres, DPM, Rodrigues, LRM, Carvalho, LB, and Teixeira, JAC, "Characterization of galactooligosaccharides produced by b-galactosidase immobilized onto magnetized Dacron". Int. Dairy J., 21: 172 (2011).

30. Neves, CMB, Simoes, MMQ, Santos, I, Domingues, FMJ, Neves, M, Paz, FAA, Silva, AMS, and Cavaleiro, JAS, "Oxidation of caffeine with hydrogen peroxide catalyzed by metalloporphyrins". Tetrahedron Lett., 52: 2898 (2011).

31. Seca, AML and Silva, AMS, "A new 4 ',7-epoxy-8,3 '-oxyneolignan from the acetone extract of Juniperus brevifolia leaves". Phytochem. Lett., 3: 126 (2010).

32. Seixas, R, Silva, AMS, and Cavaleiro, JAS, "New Synthesis of (Z)- and (E)-3-Styryl-4-quinolones". Synlett: 2257 (2010).

33. Tome, AC, Silva, AMS, Alkorta, I, and Elguero, J, "Atropisomerism and conformational aspects of meso-tetraarylporphyrins and related compounds". J. Porphyr. Phthalocyanines, 15: 1 (2011).

34. Duarte, IF, Legido-Quigley, C, Parker, DA, Swann, JR, Spraul, M, Braumann, U, Gil, AM, Holmes, E, Nicholson, JK, Murphy, GM, Vilca-Melendez, H, Heatond, N, and Lindon, JC, "Identification of metabolites in human hepatic bile using 800 MHz 1H NMR spectroscopy, HPLC-NMR/MS and UPLC-MS". Molecular Biosyst., 5: 180 (2009).

35. Graca, G, Duarte, IF, Barros, AS, Goodfellow, BJ, Diaz, S, Carreira, IM, Couceiro, AB, Galhano, E, and Gil, AM, "1H NMR Based Metabonomics of Human Amniotic Fluid for the Metabolic Characterization of Fetus Malformations". J. Proteome Res., 8: 4144 (2009).

36. Graca, G, Duarte, IF, Barros, AS, Goodfellow, BJ, Diaz, SO, Pinto, J, Carreira, IM, Galhano, E, Pita, C, and Gil, AM, "Impact of Prenatal Disorders on the Metabolic Profile of Second Trimester Amniotic Fluid: A Nuclear Magnetic Resonance Metabonomic Study". J. Proteome Res., 9: 6016 (2010).

37. Rocha, CM, Barros, AS, Gil, AM, Goodfellow, BJ, Humpfer, E, Spraul, M, Carreira, IM, Melo, JB, Bernardo, J, Gomes, A, Sousa, V, Carvalho, L, and Duarte, IF, "Metabolic Profiling of Human Lung Cancer Tissue by 1H High Resolution Magic Angle Spinning (HRMAS) NMR Spectroscopy". J. Proteome Res., 9: 319 (2010).

38. Carrola, J, Rocha, CM, Barros, AS, Gil, AM, Goodfellow, BJ, Carreira, IM, Bernardo, J, Gomes, A, Sousa, V, Carvalho, L, and Duarte, IF, "Metabolic Signatures of Lung Cancer in Biofluids: NMR-Based Metabonomics of Urine". J. Proteome Res., 10: 221 (2011).

39. Diaz, SO, Pinto, J, Graca, G, Duarte, IF, Barros, AS, Galhano, E, Pita, C, Almeida, MD, Goodfellow, BJ, Carreira, IM, and Gil, AM, "Metabolic Biomarkers of Prenatal Disorders: An Exploratory NMR Metabonomics Study of Second Trimester Maternal Urine and Blood Plasma". J. Proteome Res., 10: 3732 (2011).

40. Daniel-da-Silva, AL, Fateixa, S, Guiomar, AJ, Costa, BFO, Silva, NJO, Trindade, T, Goodfellow, BJ, and Gil, AM, "Biofunctionalized magnetic hydrogel nanospheres of magnetite and k-carrageenan". Nanotechnology, 20: 355602(2009).

41. Daniel-da-Silva, AL, Ferreira, L, Gil, AM, and Trindade, T, "Synthesis and swelling behavior of temperature responsive k-carrageenan nanogels". J. Colloid Interface Sci., 355: 512 (2011).

42. Duarte, IF, Marques, J, Ladeirinha, AF, Rocha, C, Lamego, I, Calheiros, R, Silva, TM, Marques, MPM, Melo, JB, Carreira, IM, and Gil, AM, "Analytical Approaches toward Successful Human Cell Metabolome Studies by NMR Spectroscopy". Anal. Chem., 81: 5023 (2009).

43. Duarte, IF, Lamego, I, Rocha, C, and Gil, AM, "NMR metabonomics for mammalian cell metabolism studies". Bioanalysis, 1: 1597 (2009).

44. Duarte, IF, Lamego, I, Marques, J, Marques, MPM, Blaise, BJ, and Gil, AM, "Nuclear Magnetic Resonance (NMR) Study of the Effect of Cisplatin on the Metabolic Profile of MG-63 Osteosarcoma Cells". J. Proteome Res., 9: 5877 (2010).

45. Rodrigues, JEA, Erny, GL, Barros, AS, Esteves, VI, Brandao, T, Ferreira, AA, Cabrita, E, and Gil, AM, "Quantification of organic acids in beer by nuclear magnetic resonance (NMR)-based methods". Anal. Chim. Acta, 674: 166 (2010).

46. Rodrigues, JA, Barros, AS, Carvalho, B, Brandao, T, Gil, AM, and Ferreira, ACS, "Evaluation of beer deterioration by gas chromatography-mass spectrometry/multivariate analysis: A rapid tool for assessing beer composition". J. Chromatogr. A, 1218: 990 (2011).

47. Rodrigues, JA, Barros, AS, Carvalho, B, Brandao, T, and Gil, AM, "Probing beer aging chemistry by nuclear magnetic resonance and multivariate analysis". Anal. Chim. Acta, 702: 178 (2011).

48. Jarvinen, R, Silvestre, AJD, Gil, AM, and Kallio, H, "Solid state 13C CP-MAS NMR and FT-IR spectroscopic analysis of cuticular fractions of berries and suberized membranes of potato". J. Food Compos. Anal., 24: 334 (2011).

49. Jarvinen, R, Silvestre, AJD, Holopainen, U, Kaimainen, M, Nyyssola, A, Gil, AM, Neto, CP, Lehtinen, P, Buchert, J, and Kallio, H, "Suberin of Potato (Solanum tuberosum Var. Nikola): Comparison of the Effect of Cutinase CcCut1 with Chemical Depolymerization". J. Agric. Food Chem., 57: 9016 (2009).

50. Webber, AL, Elena, B, Griffin, JM, Yates, JR, Pham, TN, Mauri, F, Pickard, CJ, Gil, AM, Stein, R, Lesage, A, Emsley, L, and Brown, SP, "Complete 1H resonance assignment of b-maltose from 1H-1H DQ-SQ CRAMPS and 1H (DQ-DUMBO)-13C SQ refocused INEPT 2D solid-state NMR spectra and first principles GIPAW calculations". Phys. Chem. Chem. Phys., 12: 6970 (2010).

51. Goodfellow, BJ, Dias, JS, Ferreira, GC, Henklein, P, Wray, V, and Macedo, AL, "The solution structure and heme binding of the presequence of murine 5-aminolevulinate synthase". Febs Lett., 505: 325 (2001).

52. Dias, JS, Macedo, AL, Ferreira, GC, Peterson, FC, Volkman, BF, and Goodfellow, BJ, "The first structure from the SOUL/HBP family of heme-binding proteins, murine P22HBP". J. Biol. Chem., 281: 31553 (2006).

53. Freire, F, Romao, MJ, Macedo, AL, Aveiro, SS, Goodfellow, BJ, and Carvalho, AL, "Preliminary structural characterization of human SOUL, a haem-binding protein". Acta Crystallogr. E, 65: 723 (2009).

54. Goodfellow, BJ, Nunes, SG, Rusnak, F, Moura, I, Ascenso, C, Moura, JJG, Volkman, BF, and Markley, JL, "Zinc-substituted Desulfovibrio gigas desulforedoxins: Resolving subunit degeneracy with nonsymmetric pseudocontact shifts". Protein Sci., 11: 2464 (2002).

55. Goodfellow, BJ, Duarte, ICN, Macedo, AL, Volkman, BF, Nunes, SG, Moura, I, Markley, JL, and Moura, JJG, "An NMR structural study of nickel-substituted rubredoxin". J. Biol. Inorg. Chem., 15: 409 (2010).

56. Duarte, IF, "Following dynamic biological processes through NMR-based metabonomics: A new tool in nanomedicine?" J. Control. Release, 153: 34 (2011).

57. Rocha, CM, Carrola, J, Barros, AS, Gil, AM, Goodfellow, BJ, Carreira, IM, Bernardo, J, Gomes, A, Sousa, V, Carvalho, L, and Duarte, IF, "Metabolic Signatures of Lung Cancer in Biofluids: NMR-Based Metabonomics of Blood Plasma". J. Proteome Res., 10: 4314 (2011).