Maria Harkiolaki

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Maria Harkiolaki is the Principal Beamline Scientist for beamline B24. Maria joined Diamond in 2013 after working at the University of Oxford.

Email: maria.harkiolaki@diamond.ac.uk
Tel: +44 (0) 1235 778757

Research Summary

  1. Research
  2. Collaborations
  3. Publications
  4. Biography
Research -

 

Structural immunology, innate and adaptive immunity, pathogen recognition & adaptation.

My primary research work focuses on pathogen recognition in the honey bee and aims to elucidate the atomic structure, biological function, field variation and temporal cellular localisation of proteins implicated in the Toll/Dif and Imd/Relish signalling pathways. All main players of pathogen recognition (toll, GNBP, PGRP, Spätzle, Serpins and Serine Proteases) are used and a full panel of ecto-domains and component sub-domains are cloned to produce soluble constructs for crystallisation screening. Diffraction-quality crystals of these are used to collect X-ray data and generate atomic resolution molecular structures. In parallel, all constructs are characterised in vitro using established binding assays and pull-down experiments and in vivo through rescue experiments in transgenic D. melanogaster. Using Xray microscopy, primary cell cultures can be assessed for their morphological properties as well as the distribution of key molecular players upon antigenic challenge.
 
HIV research - Insect immunity models - Livestock immunity - Molecular modelling.
 
Beamline components design, upgrades and maintenance.  Technique development.  User Support.
Collaborations - +

 

  • Collaborator is Dr P Ligoxygakis at the University of Oxford - Biochemistry to study Insect Innate Immunity. My role involved protein construct design and structure determination and cellular localisation. 


     
  • Collaborator is Dr A Iversen at the University of Oxford - Division of Clinical Neurology to study Immune responses to HIV infection My role is the molecular structure determination of MHC class I complexes.

Publications - +
  • F. Vaz, I. Kounatidis, G. Covas, R. M. Parton, M. Harkiolaki, I. Davis, S. R. Filipe, P. Ligoxygakis. Accessibility to Peptidoglycan Is Important for the Recognition of Gram-Positive Bacteria in Drosophila. Cell Rep. (2019) 27:2480–2492.

  • M. C. Spink, M. C. Darrow, H. Fisher, A. Burt, K. Marshall, I. Luengo, M. Harkiolaki, and L. Duke. Correlation of Cryo Soft X-ray Tomography with Cryo Fluorescence Microscopy to Characterise Cellular Organelles at Beamline B24, Diamond Light Source. Microscopy & Microanalysis (2018) 24: 374-375.

  • M. Harkiolaki, M. C. Darrow, M. C. Spink, E. Kosior, K. Dent, E. Duke. Cryo-soft X-ray tomography: Using soft X-rays to explore the ultrastructure of whole cells. Emerg. Top. Life Sci. (2018) 2: 81–92.

  • M. C. Darrow, M. Harkiolaki, M. Spink, I. Luengo, M. Basham and L. Duke. Cryo Soft X-ray Tomography and Other Techniques at Diamond Light Source. Microscopy & Microanalysis (2017) 23: 972-973.

  • V. L. Hale, J. M. Watermeyer, F. Hackett, G. Vizcay-Barrena, C. Van Ooij, J. A. Thomas, M. C. Spink, M. Harkiolaki, E. Duke, R. A. Fleck, M. J. Blackman and H. R. Saibil. Parasitophorous vacuole poration precedes its rupture and rapid host erythrocyte cytoskeleton collapse in Plasmodium falciparum egress. Proceedings of the National Academy of Sciences (2017) 114: 3439-3444.

  • J. García-Nafría, M. Fogg, M. Harkiolaki, R. Persson, K Wilson. Novel features in Bacillus subtilis dUTPases. The Febs Journal (2011) 278, 456-457.

  • J. Garcia-Nafria, M. Harkiolaki, R. Persson, M. J. Fogg and K. S. Wilson. The structure of Bacillus subtilis SPβ prophage dUTPase and its complexes with two nucleotides. Acta Cryst D (2011) 67: 167-75.

  • I. K. Macdonald*, M. Harkiolaki*, L. Hunt, W. I. Morrison, T. Connelley, S. P. Graham, E. Y. Jones, D. R. Flower and S. A. Ellis. Peptide presentation by a cattle MHC class I: Crystal structure of BoLA N*01301 bound to an immunodominant 11mer Theileria parva epitope. PLoS Pathogens (2010) 6: e1001149. *equal first authors.

  • M. Harkiolaki, T. Tsirka, M. Lewitzky, P. C. Simister, D. Joshi, L. E. Bird, E. Y. Jones, N. O’Reilly and S. M. Feller. Distinct binding modes of two epitopes in Gab2 that interact with the SH3 domain of Grb2. Structure (2009) 17: 809-22.

  • M. Harkiolaki, S. L. Holmes, P. Svendsen, J. W. Gregersen, L. T. Jensen, R. McMahon, M. A. Friese, G. van Boxel, R. Etzensperger, J. S. Tzartos, K. Kranc, S. Sainsbury, K. Harlos, E. D. Mellins, J. Palace, M. M. Esiri, P. A. van der Merwe, E. Y. Jones and L. Fugger. T Cell-mediated Autoimmune disease Due to Low-Affinity Crossreactivity to Common Microbial Peptides. Immunity (2009) 30:348-57.

  • M. Harkiolaki, R. J. Gilbert, E. Y. Jones and S. M. Feller. The C-terminal SH3 domain of CRKL as a dynamic dimerization module transiently exposing a nuclear export signal. Structure (2006) 14:1741-53.

  • C. S. Hourigan, M. Harkiolaki, N. A. Peterson, J. I. Bell, E. Y. Jones, C. A. O'Callaghan. The structure of the human allo-ligand HLA-B*3501 in complex with a cytochrome p450 peptide: steric hindrance influences TCR allo-recognition. Eur J Immunol. (2006) 36:3288-93.

  • O. V. Moroz, M. Harkiolaki, M. Y. Galperin, A. A. Vagin, D. Gonzalez-Pacanowska and K. S. Wilson. The crystal structure of a complex of Campylobacter jejuni dUTPase with substrate analogue sheds light on the mechanism and suggests the "basic module" for dimeric d(C/U)TPases. J. Mol. Biol. (2004) 342:1583-1597.

  • M. Lewitzky*, M. Harkiolaki*, M. C. Domart, E. Y. Jones and S. M. Feller. Mona/Gads SH3C binding to hematopoietic progenitor kinase 1 (HPK1) combines an atypical SH3 binding motif, R/KXXK, with a classical PXXP motif embedded in a polyproline type II (PPII) helix. J. Biol. Chem. (2004) 279: 28724-28732. * Joint first authors.

  • M. Harkiolaki, E. J. Dodson, V. Bernier-Villamor, J. P. Turkenburg, D. González-Pacanowska, and K. S. Wilson. The crystal structure of Trypanosoma cruzi dUTPase reveals a novel dUTP/dUDP binding fold. Structure (2004) 12: 41-53.

  • M. Harkiolaki, M. Lewitzky, R. J.C. Gilbert, E.Y. Jones, R. P. Bourette, G. Mouchiroud, H. Sondermann, I.l Moarefi, and S. M. Feller. Structural basis for SH3 domain-mediated high-affinity binding between Mona/Gads and SLP-76. EMBO J. (2003) 22: 2571 - 2582.

  • J. Brown, T. S. Walter, L. Carter, N. G. A. Abrescia, A. R. Aricescu, T. D. Batuwangala, L. E. Bird, N. Brown, P. P. Chamberlain, S. J. Davis, E. Dubinina, J. Endicott, J. A. Fennelly, R. J. C. Gilbert, M. Harkiolaki, W.-C. Hon, F. Kimberley, C. A. Love, E. J. Mancini, R. Manso-Sancho, C. E. Nichols, R. A. Robinson, G. C. Sutton, N. Schueller, M. C. Sleeman, G. B. Stewart-Jones, M. Vuong, J. Welburn, Z. Zhang, D. K. Stammers, R. J. Owens, E. Y. Jones, K. Harlos and D. I. Stuart. A procedure for setting up high-throughput nanolitre crystallization experiments. II. Crystallization results. J. Appl. Cryst. (2003) 36, 315-318.

  • A. Perrakis, M. Harkiolaki, K. S. Wilson and V. S. Lamzin. ARP/wARP and molecular replacement. Acta Cryst. (2001) D57, 1445-1450.

  • M. Harkiolaki, A. M. Brzozowski , D. Gonzalez-Pacanowska, F. Hidalgo-Zarco and K. S. Wilson. New crystal forms of Trypanosoma cruzi dUTPase. Acta Cryst. (2001) D57, 915-917.

  • R. Persson*, M. Harkiolaki*, J. McGeehan and K. S. Wilson. Crystallization and preliminary crystallographic analysis of deoxyuridine 5'-triphosphate nucleotidohydrolase from Bacillus subtilis. Acta Cryst. (2001) D57, 876-878. * Joint first authors.

Biography - +

 

Dr Harkiolaki has been at Diamond since 2012. She has been involved with the design, procurement and installation of B24 components as well as the development of other high resolution microscopy methods that will be used in a correlative fashion to the data generated by the B24 full field X-ray microscope. Previously, she was a Principal Investigator at the Structural Biology Laboratory at the Nuffield Department of Clinical Medicine of the University of Oxford. While there she worked on a range of biomedical research projects with emphasis initially on human adaptive immunity, later expanding to model studies of innate immunity. In specific, she studied MHC class II molecules relating to multiple sclerosis, MHC class I molecules relating to allo reactivity & HIV response as well as MHC class I complexes expressed upon antigenic challenge in livestock. Moreover, she has developed a library of innate immunity molecules relating to two species of honey bee: western and asiatic, to be studied further and in conjunction with emerging data from other laboratories. Dr Harkiolaki studied for her PhD at the University of York, under the supervision of Professors K. Wilson and E. Dodson.

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