Biotechnology

Microbial and Molecular Biotechnology

Host-pathogen Interactions

Rickettsia

Proteolysis

Aspartic Proteases

Biotechnology

Recombinant Protein Production


Research lines

Understand the role of proteolysis in the context of infection, using spotted fever group rickettsiae as our working model, to identify proteases/proteolytic machineries as novel biological targets

Understand the role of macrophage permissiveness in rickettsial pathogenesis and decipher how mildly pathogenic SFG Rickettsia target and manipulate host cell metabolic and proteostatic responses

Decipher the role of cell-to-cell communication for the development of rickettsial infections

Develop new biotech applications based on proteases and proteases’ engineering"

Overview

The activities of the Microbial and Molecular Biotechnology Group are focused on performing high-quality research and advanced training to obtain a better understanding of the mechanisms of disease and pathogenicity in obligate intracellular pathogens, as is the case of many Rickettsia spp. With our work, we expect to contribute to the identification of new factors and/or molecular pathways that may constitute pathogen- or host-directed targets for therapeutic intervention or the development of new diagnostics.

Our research is grounded on interdisciplinary collaborations with an international orientation. We have made significant contributions both in fundamental aspects of structure-function relationships of aspartic proteases as well as in exploring their biotechnological potential. Also, we have been contributing to establish macrophage colonization as a new virulence trait in Rickettsia. Therefore, allying our expertise in the proteolysis field with our experience with SFG Rickettsia, our current research program aims at exploring two understudied aspects of rickettsial biology:

  1. How proteolytic pathways modulate Rickettsia-host cell interactions;
  2. How mildly pathogenic Rickettsia hijack host cell function;

Our research programs combine methodologies from cell biology, molecular biology, recombinant DNA technology, heterologous protein production, protein chemistry, enzymology, complemented with various system-wide quantitative proteomics approaches. We are a BSL-2 lab, with state of the art equipment for: Cell culture and handling of pathogenic organisms (biological safety cabinets, centrifuge safety containers); Protein expression (incubators up to 12L, high capacity centrifuges; High-Pressure homogenizer); Protein Purification (low and medium pressure chromatography, Tangential Filtration systems); Protein Characterization (HPLC, Fluorimeter, Spectrophotometer); and Plant growth (FitoClima PLH 12000 ‘walk-in’);

Publications

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Information about journal articles, updated at 05-12-2021, from platform CIÊNCIAVITAE.

Biochemical diversity of cardoon flowers (Cynara cardunculus L.): Predicting PDO Mediterranean cheese textures

Barracosa, P.; Simões, I.; Martins, A.P.; Barros, M.; Pires, E., 2021. Food Bioscience. 39. 2021. http://dx.doi.org/10.1016/j.fbio.2020.100805 . 10.1016/j.fbio.2020.100805 . published Food Bioscience

Macrophages Infected by a Pathogen and a Non-pathogen Spotted Fever Group Rickettsia Reveal Differential Reprogramming Signatures Early in Infection

Curto, Pedro; Riley, Sean P.; Simões, Isaura; Martinez, Juan J., 2019. Frontiers in Cellular and Infection Microbiology. 9. 2019. http://dx.doi.org/10.3389/fcimb.2019.00097 . 10.3389/fcimb.2019.00097 . Frontiers in Cellular and Infection Microbiology

A Pathogen and a Non-pathogen Spotted Fever Group Rickettsia Trigger Differential Proteome Signatures in Macrophages

Curto, Pedro; Santa, Cátia; Allen, Paige; Manadas, Bruno; Simões, Isaura; Martinez, Juan J., 2019. Frontiers in Cellular and Infection Microbiology. 9. 2019. http://dx.doi.org/10.3389/fcimb.2019.00043 . 10.3389/fcimb.2019.00043 . Frontiers in Cellular and Infection Microbiology

An atypical aspartic protease modulates lateral root development in Arabidopsis thaliana

Soares, André; Niedermaier, Stefan; Faro, Rosário; Loos, Andreas; Manadas, Bruno; Faro, Carlos; Huesgen, Pitter F; Cheung, Alice Y; Simões, Isaura, 2019. Journal of Experimental Botany. 2157 - 2171. 7. 70. 2019. http://dx.doi.org/10.1093/jxb/erz059 . 10.1093/jxb/erz059 . Journal of Experimental Botany

Atypical and nucellin-like aspartic proteases: emerging players in plant developmental processes and stress responses

André Soares; Sofia M Ribeiro Carlton; Isaura Simões, 2019. Journal of Experimental Botany. 2019. https://doi.org/10.1093/jxb/erz034 . 10.1093/jxb/erz034 . Journal of Experimental Botany

Use of recombinant proteins as a simple and robust normalization method for untargeted proteomics screening: exhaustive performance assessment

Anjo, Sandra Isabel; Simões, Isaura; Castanheira, Pedro; Grãos, Mário; Manadas, Bruno, 2019. Talanta. 205. 2019. http://dx.doi.org/10.1016/j.talanta.2019.120163 . 10.1016/j.talanta.2019.120163 . Talanta

Cardoon-based rennets for cheese production

Almeida, Carla Malaquias; Simões, Isaura, 2018. Applied Microbiology and Biotechnology. 4675 - 4686. 11. 102. 2018. http://dx.doi.org/10.1007/s00253-018-9032-3 . 10.1007/s00253-018-9032-3 . Applied Microbiology and Biotechnology

Functional and structural characterization of synthetic cardosin B-derived rennet

Almeida, Carla Malaquias; Manso, José A.; Figueiredo, Ana C.; Antunes, Liliana; Cruz, Rui; Manadas, Bruno; Bur, Daniel; et al, 2017. Applied Microbiology and Biotechnology. 6951 - 6968. 18. 101. 2017. http://dx.doi.org/10.1007/s00253-017-8445-8 . 10.1007/s00253-017-8445-8 . Applied Microbiology and Biotechnology

Differences in Intracellular Fate of Two Spotted Fever Group Rickettsia in Macrophage-Like Cells

Curto, Pedro; Simões, Isaura; Riley, Sean P.; Martinez, Juan J., 2016. Frontiers in Cellular and Infection Microbiology. 6. 2016. http://dx.doi.org/10.3389/fcimb.2016.00080 . 10.3389/fcimb.2016.00080 . Frontiers in Cellular and Infection Microbiology

Enzymatic properties, evidence for in vivo expression, and intracellular localization of shewasin D, the pepsin homolog from Shewanella denitrificans

Leal, Ana Rita; Cruz, Rui; Bur, Daniel; Huesgen, Pitter F.; Faro, Rosário; Manadas, Bruno; Wlodawer, Alexander; Faro, Carlos; Simões, Isaura, 2016. Scientific Reports. 1. 6. 2016. http://dx.doi.org/10.1038/srep23869 . 10.1038/srep23869 . Scientific Reports

Biophysical characterization of laforin–carbohydrate interaction

Dias, David M.; Furtado, Joana; Wasielewski, Emeric; Cruz, Rui; Costello, Bernard; Cole, Lindsay; Faria, Tiago Q.; et al, 2016. Biochemical Journal. 335 - 345. 3. 473. 2016. http://dx.doi.org/10.1042/bj20141555 . 10.1042/bj20141555 . Biochemical Journal

Structure of RC1339/APRc fromRickettsia conorii, a retropepsin-like aspartic protease

Li, Mi; Gustchina, Alla; Cruz, Rui; Simões, Marisa; Curto, Pedro; Martinez, Juan; Faro, Carlos; Simões, Isaura; Wlodawer, Alexander, 2015. Acta Crystallographica Section D Biological Crystallography. 2109 - 2118. 10. 71. 2015. http://dx.doi.org/10.1107/s1399004715013905 . 10.1107/s1399004715013905 . Acta Crystallographica Section D Biological Crystallography

RC1339/APRc from Rickettsia conorii Is a Novel Aspartic Protease with Properties of Retropepsin-Like Enzymes

Cruz, Rui; Huesgen, Pitter; Riley, Sean P.; Wlodawer, Alexander; Faro, Carlos; Overall, Christopher M.; Martinez, Juan J.; Simões, Isaura, 2014. PLoS Pathogens. 8. 10. 2014. http://dx.doi.org/10.1371/journal.ppat.1004324 . 10.1371/journal.ppat.1004324 . PLoS Pathogens

Engineering a cardosin B-derived rennet for sheep and goat cheese manufacture

Almeida, Carla Malaquias; Gomes, David; Faro, Carlos; Simões, Isaura, 2014. Applied Microbiology and Biotechnology. 269 - 281. 1. 99. 2014. http://dx.doi.org/10.1007/s00253-014-5902-5 . 10.1007/s00253-014-5902-5 . Applied Microbiology and Biotechnology

Establishing the Yeast Kluyveromyces lactis as an Expression Host for Production of the Saposin-Like Domain of the Aspartic Protease Cirsin

Curto, Pedro; Lufrano, Daniela; Pinto, Cátia; Custódio, Valéria; Gomes, Ana Catarina; Trejo, Sebastián A.; Bakás, Laura; et al, 2013. Applied and Environmental Microbiology. 86 - 96. 1. 80. 2013. http://dx.doi.org/10.1128/aem.03151-13 . 10.1128/aem.03151-13 . Applied and Environmental Microbiology

Effect of a-Synuclein on Amyloid ß-Induced Toxicity: Relevance to Lewy Body Variant of Alzheimer Disease

Resende, Rosa; Marques, Sueli C. F.; Ferreiro, Elisabete; Simões, Isaura; Oliveira, Catarina R.; Pereira, Cláudia M. F., 2013. Neurochemical Research. 797 - 806. 4. 38. 2013. http://dx.doi.org/10.1007/s11064-013-0982-7 . 10.1007/s11064-013-0982-7 . Neurochemical Research

Molecular cloning and characterization of procirsin, an active aspartic protease precursor from Cirsium vulgare (Asteraceae)

Lufrano, Daniela; Faro, Rosário; Castanheira, Pedro; Parisi, Gustavo; Veríssimo, Paula; Vairo-Cavalli, Sandra; Simões, Isaura; Faro, Carlos, 2012. Phytochemistry. 7 - 18. 81. 2012. http://dx.doi.org/10.1016/j.phytochem.2012.05.028 . 10.1016/j.phytochem.2012.05.028 . Phytochemistry

Chlapsin, a chloroplastidial aspartic proteinase from the green algae Chlamydomonas reinhardtii

Almeida, Carla Malaquias; Pereira, Cláudia; da Costa, Diana Soares; Pereira, Susana; Pissarra, José; Simões, Isaura; Faro, Carlos, 2012. Planta. 283 - 296. 1. 236. 2012. http://dx.doi.org/10.1007/s00425-012-1605-2 . 10.1007/s00425-012-1605-2 . Planta

Shewasin A, an active pepsin homolog from the bacterium Shewanella amazonensis

Simões, Isaura; Faro, Rosário; Bur, Daniel; Kay, John; Faro, Carlos, 2011. FEBS Journal. 3177 - 3186. 17. 278. 2011. http://dx.doi.org/10.1111/j.1742-4658.2011.08243.x . 10.1111/j.1742-4658.2011.08243.x . FEBS Journal

Characterization of Recombinant CDR1, anArabidopsisAspartic Proteinase Involved in Disease Resistance

Simões, Isaura; Faro, Rosário; Bur, Daniel; Faro, Carlos, 2007. Journal of Biological Chemistry. 31358 - 31365. 43. 282. 2007. http://dx.doi.org/10.1074/jbc.m702477200 . 10.1074/jbc.m702477200 . Journal of Biological Chemistry

Molecular analysis of the interaction between cardosin A and phospholipase Da

Simões, Isaura; Mueller, Eva-Christina; Otto, Albrecht; Bur, Daniel; Cheung, Alice Y.; Faro, Carlos; Pires, Euclides, 2005. FEBS Journal. 5786 - 5798. 22. 272. 2005. http://dx.doi.org/10.1111/j.1742-4658.2005.04967.x . 10.1111/j.1742-4658.2005.04967.x . FEBS Journal

Structure and function of plant aspartic proteinases

Simões, Isaura; Faro, Carlos, 2004. European Journal of Biochemistry. 2067 - 2075. 11. 271. 2004. http://dx.doi.org/10.1111/j.1432-1033.2004.04136.x . 10.1111/j.1432-1033.2004.04136.x . European Journal of Biochemistry

Cloning and Characterization of cDNA Encoding Cardosin A, an RGD-containing Plant Aspartic Proteinase

Faro, Carlos; Ramalho-Santos, Miguel; Vieira, Margarida; Mendes, Alexandra; Simões, Isaura; Andrade, Rita; Veri´ssimo, Paula; et al, 1999. Journal of Biological Chemistry. 28724 - 28729. 40. 274. 1999. http://dx.doi.org/10.1074/jbc.274.40.28724 . 10.1074/jbc.274.40.28724 . Journal of Biological Chemistry

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