Host-pathogen Interactions
Rickettsia
Proteolysis
Aspartic Proteases
Biotechnology
Recombinant Protein Production
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"
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:
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’);
Information about journal articles, updated at 08-12-2024, from platform CIÊNCIAVITAE.
Cx43-mediated sorting of miRNAs into extracellular vesicles
Martins-Marques, Tania; Costa, Marina C; Catarino, Steve; Simoes, Isaura; Aasen, Trond; Enguita, Francisco J; Girao, Henrique, 2022. EMBO reports. 2022. http://dx.doi.org/10.15252/embr.202154312 . 10.15252/embr.202154312 . published EMBO reports
Moonlighting in Rickettsiales: Expanding Virulence Landscape
Matos, Ana Luísa; Curto, Pedro; Simões, Isaura, 2022. Tropical Medicine and Infectious Disease. 2. 7. 2022. http://dx.doi.org/10.3390/tropicalmed7020032 . 10.3390/tropicalmed7020032 . Tropical Medicine and Infectious Disease
The Retropepsin-Type Protease APRc as a Novel Ig-Binding Protein and Moonlighting Immune Evasion Factor of Rickettsia
Curto, Pedro; Barro, Andreia; Almeida, Carla; Vieira-Pires, Ricardo S.; Simões, Isaura, 2021. mBio. 6. 12. 2021. http://dx.doi.org/10.1128/mbio.03059-21 . 10.1128/mbio.03059-21 . mBio
Spotted Fever Group Rickettsia Trigger Species-Specific Alterations in Macrophage Proteome Signatures with Different Impacts in Host Innate Inflammatory Responses
Curto, Pedro; Santa, Cátia; Cortes, Luísa; Manadas, Bruno; Simões, Isaura, 2021. Microbiology Spectrum. 3. 9. 2021. http://dx.doi.org/10.1128/spectrum.00814-21 . 10.1128/spectrum.00814-21 . Microbiology Spectrum
Introduction of a Glycine Linker Connecting the Heavy and Light Chains in Synthetic Cardosin B-Derived Rennet Changes the Specificity of Subpocket S3'
Figueiredo, Pedro R.; Santos, Sónia F. G.; Almeida, Beatriz C.; Simões, Isaura; Carvalho, Alexandra T. P., 2021. The Journal of Physical Chemistry B. 4368 - 4374. 17. 125. 2021. http://dx.doi.org/10.1021/acs.jpcb.1c01826 . 10.1021/acs.jpcb.1c01826 . The Journal of Physical Chemistry B
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 . Food Bioscience
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
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
Soares, André; Ribeiro Carlton, Sofia M; Simões, Isaura, 2019. Journal of Experimental Botany. 2059 - 2076. 7. 70. 2019. http://dx.doi.org/10.1093/jxb/erz034 . 10.1093/jxb/erz034 . Journal of Experimental Botany
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
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
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
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
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
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, 2014. Applied and Environmental Microbiology. 86 - 96. 1. 80. 2014. 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, an Arabidopsis Aspartic 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
5 28 out of 28 Publications
The role of cell-to-cell communication in rickettsial pathogenesis: an approach to unravel new virulence factors
2023-03-01
Hijack the host or get killed: delineating Rickettsia survival strategies for development of targeted therapeutic strategies
2018-07-26
Deciphering the Rickettsia toolbox to hijack the host
2020-04-01
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