Dr Paula Salgado
Lecturer in Macromolecular Crystallography
- Email: email@example.com
- Telephone: 01912087432
- Personal Website: http://sbl.ncl.ac.uk/people/paula_research.shtml
- Address: Institute for Cell and Molecular Biosciences
Faculty of Medical Sciences
3rd Floor Cookson Building
Newcastle upon Tyne, NE2 4HH, UK
2012 - present Lecturer of MacromolecularCrystallography, ICaMB, Newcastle University
2011 -2012 Research Associate, Division of Cell and Molecular Biology, Department of Life Sciences, Imperial College London, UK
2008 – 2011 Research Associate, Division of Molecular Biosciences, Department of Life Sciences, Imperial College London, UK
2005 – 2008 Research Associate, School of Crystallography, Birkbeck College/University College, London, UK
2001 – 2005 PhD in Structural Biology, Henry Wellcome Trust Building for Human Genetics, University of Oxford and Open University. "Structural Studies of RNA-dependent RNA polymerases" Supervisors: Dr Jon Grimes and Prof Dave I Stuart
1993 -1999 BSc in Biochemistry, Science Faculty & Institute of Biomedical Sciences “Abel Salazar”, University of Porto, Portugal. Research Project: "Structure determination of transthyretin variants"
November 2014 - November 2017 - New Investigator Research Grant, MRC
April 2013 - March 2014 - Royal Society Research Grant
March - June 2011 - Wellcome Trust Value In People Award
Oct 01 - Oct 2005 - PhD Fellowship (part of Human Frontier Science Program Research Grant)
As a Structural Biologist, my main research interest is in the determination and understanding of the structure of proteins associated to human disease.
I am particularly interested in proteins involved in disease caused by infectious agents, aiming at providing molecular details of key mechanisms of host-pathogen interactions, relevant for the development of more effective treatments.
Current focus in the lab is on proteins from two human pathogens particularly relevant in hospital environment: the bacteria Clostridium difficile and the fungus Candida albicans.
Clostridium difficile S-layer
C. difficile is now the most prevalent hospital acquired infection in the UK. These gram-positive bacteria have an outside para-crystalline layer (S-layer) that is presumed to act as a protective shield and that is thought to be implicated in virulence.
In collaboration with Prof Neil Fairweather at Imperial College London, we focus on understanding S-layer organisation, primarily by determining the structure of the major S-layer protein constituent, SlpA. We are also studying other cell wall proteins identified in C. difficile, aiming to understand their macromolecular details with our structural studies that complement work developed at Imperial College.
Biofilms are microbial masses that adhere to biological or non-biological surfaces and have been implicated in both the spread and onset of infections. Importantly, they provide a protective environment as microorganisms in biofilms are more resistant to antibiotics and host defences.
Candida albicans can cause a wide range of diseases and the formation of biofilms on catheters and medical implants is a significant source of systemic infections in the hospital environment.
The CFEM-containing family of proteins, characterised by an eight-cysteine motif unique to fungal species, has been implicated in biofilm formation and could provide a structural scaffold for new rational drug development. Our research, in collaboration with Dr Carol Munro at the University of Aberdeen, is focusing on providing details into the structural organisation of this unusual motif and understand its role in candidal infections.
Recently, while working at Imperial College London with Dr Ernesto Cota, I determined the structure of the first candidal adhesin, Als9. The structural model allowed the identification of a flexible unusual binding cleft in these adhesins that accounted for the observed relatively unspecific ligand binding nature of the Als family.
RNA-dependent RNA polymerases (RdRPs)
During my PhD, I determined structural models of several complexes of an RdRP of a dsRNA virus (bacteriophage Phi6) with template RNA and different cations, contributing to the elucidation of the mechanisms of initiation and inhibition of RNA polymerisation. This viral RdRP is a close structural homologue of the Hepatitis C virus polymerase and is therefore extensively used as a model enzyme. Structural information derived from the bacteriophage protein has allowed a better understanding of the replication mechanisms associated with the human pathogen, hence contributing to rational design and development of more effective treatments.
I also determined the structure of an RdRP involved in RNA silencing, the first such cellular polymerase model to be described. Surprisingly, the model revealed a catalytic site reminiscent of that found in the multi-complex enzymes involved in DNA transcription (DNA-dependent RNA polymerases, DdRPs). These observations prompted a structural-based evolutionary analysis of polymerases that highlighted unexpected links between RNA silencing and DNA transcription mechanisms.
- Dembek M, Willing SE, Hong HA, Hosseini S, Salgado PS, Cutting SM. Inducible expression of spo0A as a universal tool for studying sporulation in Clostridium difficile. Frontiers in Microbiology 2017, 8, 1793.
- Serrano M, Crawshaw AD, Dembek M, Monteiro JM, Pereria FC, Gomes de Pinho M, Fairweather NF, Salgado PS, Henriques AO. The SpollQ-SpolllAH complex of Clostridium difficile controls forespore engulfment and late stages of gene expression and spore morphogenesis. Molecular Microbiology 2016, 100(1), 204-228.
- Warren AJ, Crawshaw AD, Trincao J, Aller P, Alcock S, Nistea I, Salgado PS, Evans G. In vacuo X-ray data collection from graphene-wrapped protein crystals. Acta Crystallographica Section D: Biological Crystallography 2015, 71, 2079-2088.
- Crawshaw AD, Serrano M, Stanley WA, Henriques AO, Salgado PS. A mother cell-to-forespore channel: current understanding and future challenges. FEMS Microbiology Letters 2014, 358(2), 129-136.
- Lin J, Soon-Hwan O, Jones R, Garnett JA, Salgado PS, Rusnakova S, Matthews S, Hoyer LL, Cota E. The Peptide-Binding Cavity is Essential for Als3-mediated Adhesion of Candida albicans to Human Cells. Journal of Biological Chemistry 2014, M114, 547877.
- Douse CH, Green JL, Salgado PS, Simpson PJ, Thomas JC, Langsley G, Holder AA, Tate EW, Cota E. Regulation of the Plasmodium motor complex: Phosphorylation of myosin A tail-interacting protein (MTIP) loosens its grip on MyoA. Journal of Biological Chemistry 2012, 287(44), 36968-36977.
- Taylor JD, Zhou Y, Salgado PS, Patwardhan A, McGuffie M, Pape T, Grabe G, Ashman E, Constable SC, Simpson PJ, Lee W, Cota E, Chapman MR, Matthews SJ. Atomic resolution Insights into Curli Fiber Biogenesis. Structure 2011, 19(9), 1307-1316.
- Salgado PS, Yan R, Rowan F, Cota E. Expression, crystallization, preliminary X-ray data analysis of NT-ALS9, a fungal adhesin from Candida albicans. Acta Crystallographica. Section F: Structural Biology and Crystallization Communications 2011, 67(4), 467-470.
- Salgado PS, Taylor JD, Cota E, Mathews SJ. Extending the usability of the phasing power of diselenide bonds: SeCys SAD phasing of CsgC using a non-auxotrophic strain. Acta Crystallographica. Section D: Biological Crystallography 2011, 67(p.1), 8-13.
- Salgado PS, Yan R, Taylor JD, Buchnell L, Jones R, Hoyer L, Matthews SJ, Simpson P, Cota E. Structural basis for the broad specificity to host-cell ligands by the pathogenic fungus Candida albicans. Proceedings of the National Academy of Sciences 2011, 108(38), 15775-15779.
- Poranen MM, Salgado PS, Koivunen MRL, Wright S, Bamford DH, Stuart DI, Grimes JM. Structural explanation for the role of Mn2+ in the activity of Φ6 RNA-dependent RNA polymerase. Nucleic Acids Research 2008, 36(20), 6633-6644.
- Salgado PS, Laurila MRL, Makeyev EV, Bamford DH, Stuart DI, Grimes JM. The structure of an RNAi polymerase links RNA silencing and transcription. PLoS Biology 2006, 4(12), e434.
- Laurila MRL, Salgado PS, Stuart DI, Grimes JM, Bamford DH. Back-priming mode of Phi6 RNA-dependent RNA polymerase. Journal of General Virology 2005, 86(2).
- Laurila MR, Salgado PS, Makeyev EV, Nettelship J, Stuart DI, Grimes JM, Bamford DH. Gene silencing pathway RNA-dependent RNA polymerase of Neurospora crassa: yeast expression, crystallization of selenomethionated QDE-1 protein. Journal of Structural Biology 2005, 149(1), 111-115.
- Salgado PS, Walsh MA, Laurila MRL, Stuart DI, Grimes JM. Going soft and SAD with manganese. Acta Biol Crystallographica D 2005, D61(1), 108-111.
- Salgado PS, Makeyev EV, Butcher SJ, Bamford DH, Stuart DI, Grimes JM. The structural basis for RNA specificity, Ca2+ inhibition of an RNA-dependent RNA polymerase. Structure 2004, 12(2), 307-316.