Research Lab

XTAL - Macromolecular Crystallography

xtal 2023
Research Interests

We use structural molecular biology tools to study proteins and protein-ligand complexes, using X-ray Crystallography, Cryo-Electron Microscopy, Small Angle X-ray Scattering (SAXS) and other biophysical methods.


Our main research areas are:

  1. Mechanistic studies (metalloenzymes, in particular those containing Mo, W and Fe centers, with impact on health and on the environment);
  2. Drug metabolism and ligand discovery;
  3. Glycan-protein and protein-protein interactions;
  4. Novel strategies for improved protein crystallization.
Research Highlights
Discovering 3D structures of proteins to clarify enzymatic mechanisms and understand function


a) Reduction of CO2 to formate – only few  enzymes in nature can chemically convert carbon dioxide, the greenhouse gas, into useful products. We could unveil new molecular features and add new insights to the catalytic mechanism of a tungstendependent formate dehydrogenase. This knowledge will prompt to design and analyse mechanistic relevant variants towards a more efficient enzyme.

b) Drug Metabolism – Human aldehyde oxidase (hAOX1) is an enzyme of emerging importance in phase I drug metabolism that we have extensively studied (complexes and variants). Recently we have computationaly studied its catalytic mechanism that enabled a deeper understanding of the intricacies of hAOX1 reactivity. This will help to tune drug candidates with impact in drug discovery.


highlight 2020

Representative Projects

  • “Glycan-human microbiome interaction: glycan diversity and recognition in the human gut with impact on health and nutrition”, FCT-MCTES, Total funding: €234,280, Unit funding: € 176,405 + € 15,625 (NZYTech) , Maria Angelina de Sá Palma (PI).
  • “Advances in MUC1 Glycan Cancer Antigens: From structure to function in the fight against cancer”, FCTMCTES, Total funding: €238,195, Unit funding: € 176,266 , Maria Angelina de Sá Palma (Co-PI).
  • “B.EST.CBM: an integrative structural biology approach to characterise the protein-carbohydrate microbial recognition”, FCT-MCTES, Total funding: €199,884, Unit funding: €120,684, Ana Luísa Carvalho (PI).
  • “Carbon monoxide guided shuttles (COGSs) to fight rheumatoid arthritis”, FCT-MCTES, Total funding: €199,996, Unit funding: €117,996, Teresa Santos-Silva (PI).
  • “Aldehyde oxidases in drug metabolism and biocatalysis: searching for the substrate specificity and catalytic mechanism”, FCT-MCTES, Total funding: €198,996, Unit funding: €162,792, Maria João Romão (PI).

Selected Publications

Oliveira, AR; Mota, C; Klymanska, K; Biaso, F; Romao, MJ; Guigliarelli, B; Pereira, IC. 2022. Spectroscopic and Structural Characterization of Reduced Desulfovibrio vulgaris Hildenborough W-FdhAB Reveals Stable Metal Coordination during Catalysis. ACS Chemical Biology, 17, DOI: 10.1021/acschembio.2c00336
Esteves, C; Palma, SICJ; Costa, HMA; Alves, C; Santos, GMC; Ramou, E; Carvalho, AL; Alves, V; Roque, ACA. 2022. Tackling Humidity with Designer Ionic Liquid-Based Gas Sensing Soft Materials. ADVANCED MATERIALS, DOI: 10.1002/adma.202107205
Mota, C; Diniz, A; Coelho, C; Santos-Silva, T; Esmaeeli, M; Leimkuhler, S; Cabrita, EJ; Marcelo, F; Romao, MJ. 2021. Interrogating the Inhibition Mechanisms of Human Aldehyde Oxidase by X-ray Crystallography and NMR Spectroscopy: The Raloxifene Case. JOURNAL OF MEDICINAL CHEMISTRY, 64, DOI: 10.1021/acs.jmedchem.1c01125
Bras, NF; Neves, RPP; Lopes, FAA; Correia, MAS; Palma, AS; Sousa, SF; Ramos, MJ. 2021. Combined in silico and in vitro studies to identify novel antidiabetic flavonoids targeting glycogen phosphorylase. BIOORGANIC CHEMISTRY, 108, DOI: 10.1016/j.bioorg.2020.104552
Polino, M; Rho, HS; Pina, MP; Mallada, R; Carvalho, AL; Romao, MJ; Coelhoso, I; Gardeniers, JGE; Crespo, JG; Portugal, CAM. 2021. Protein Crystallization in a Microfluidic Contactor with Nafion (R) 117 Membranes. MEMBRANES, 11, DOI: 10.3390/membranes11080549
Lima, CDL; Coelho, H; Gimeno, A; Trovao, F; Diniz, A; Dias, JS; Jimenez-Barbero, J; Corzana, F; Carvalho, AL; Cabrita, EJ; Marcelo, F. 2021. Structural Insights into the Molecular Recognition Mechanism of the Cancer and Pathogenic Epitope, LacdiNAc by Immune-Related Lectins. CHEMISTRY-A EUROPEAN JOURNAL, DOI: 10.1002/chem.202100800
Raquel dos Santos; Maria João Romão; Ana Cecília ARoque; Ana Luísa Carvalho. 2021. Magnetic particles used in a new approach for designed protein crystallization. CRYSTENGCOMM, DOI: 10.1039/d0ce01529f
Oliveira, AR; Mota, C; Mourato, C; Domingos, RM; Santos, MFA; Gesto, D; Guigliarelli, B; Santos-Silva, T; Romao, MJ; Pereira, IAC. 2020. Toward the Mechanistic Understanding of Enzymatic CO2 Reduction. ACS Catalysis, 10, DOI: 10.1021/acscatal.0c00086
Terao, M; Garattini, E; Romao, MJ; Leimkuhler, S. 2020. Evolution, expression, and substrate specificities of aldehyde oxidase enzymes in eukaryotes. JOURNAL OF BIOLOGICAL CHEMISTRY, 295, DOI: 10.1074/jbc.REV119.007741
Gomes, AS; Ramos, H; Gomes, S; Loureiro, JB; Soares, J; Barcherini, V; Monti, P; Fronza, G; Oliveira, C; Domingues, L; Bastos, M; Dourado, DFAR; Carvalho, AL; Romão, MJ; Pinheiro, B; Marcelo, F; Carvalho, A; Santos, MMM; Saraiva, L. 2020. SLMP53-1 interacts with wild-type and mutant p53 DNA-binding domain and reactivates multiple hotspot mutations. BIOCHIMICA ET BIOPHYSICA ACTA-GENERAL SUBJECTS, 1864(1), DOI: 10.1016/j.bbagen.2019.129440