The 3D structure determination of proteins using X-ray Diffraction is one of the core services of our Infrastructure. Knowledge of the 3D structure of macromolecular targets of pharmaceutical and biotechnological interest and their complexes with small molecules or other proteins may reveal the structure-function relationships that dictate their mechanism of action. Proteins participate in a plethora of metabolic pathways and controlling their interactions with their counterparts or bioactive compounds may allow the development of therapeutic interventions with significant socioeconomic impact.
Potential users: Scientists working in academic/research organizations, health sciences, hospitals, industries (pharmaceutical, food, agricultural, biofuel, paper, chemical) and SMEs.
Instruct-EL hub offers integrated structural biology services using the available infrastructure at NHRF and jointly with partner research infrastructures. It operates In close cooperation with research groups at NHRF specialized in the fields of computational chemistry, organic synthesis, biology, biotechnology and medicine. The quality of the samples prior to crystallization and exposure of the crystals to X-rays may be tested by biophysical methods.
Selected structural analysis services: X-ray diffraction data collection from protein crystals; Preliminary characterization of protein crystals; 3D structure determination of proteins; Structure-based design of potential drugs, therapeutic agents, ligands; Structural analysis from diffraction data collected at large scale facilities.
Phase identification of bulk and film samples (Pharmaceutical Industry)
Crystallite size determination
Crystallinity percentage
Solid state reactions up to 1173K
Configuration: θ-θ horizontal Bragg-Brentanno geometry
Goniometer: 600mm circle goniometer
Source: CuKa (1.6kW max)
Primary optics: fixed slits, knife edge collimator
Diffracted optics: fixed antiscatter slits, Cu absorber, soller slits, monochromator
Detectors: Scintillation, 1-D solid state super fast detector (Lynx-Eye)
Parallel beam optics: Gobel mirror
Available options: Grazing incidence XRD, Thin film measurements (XRR), high temperature furnace (RT-900oC) working under reductive and oxidative atmospheres
Visit the provider's website: http://inspired.aua.gr/instruct/
Server consists of 24 units of 8-core Intel Xeon 2.8GHz workstations with 32GB internal memory each.
The D8 Venture system includes:
IμS microfocus X-ray source, Cu radiation, air cooled
Four-circle goniometer, computer-controlled, crystal-detector distance = 35-240mm, sphere of confusion of less than 7 micrometer
Detector PHOTON 100, CMOS technology, air-cooled, 100 x 100mm active area, 240e/Cu sensitivity, shutterless data collection
800 Series Cryostream cooler, Oxford Cryosystems, AD51 Dry Air Unit, Integrated Autofill system, 60 lt and 150 lt dewar vessels.
APEX3 and PROTEUM software
Single crystal Xray diffractometer from Bruker is an X-ray generator which enables us to check crystals of various sizes and collect X-ray diffraction data from a single crystal sample. In addition to the single source configuration of the D8 QUEST, the larger enclosure of the D8 VENTURE hosts all the high intensity sources for working on even smaller, weakly diffracting crystals. The D8 VENTURE offers highest experimental flexibility, with excellent sample accessibility and visibility. It is engineered with high modularity from best-in-class components and is available in popular all-air-cooled configurations. The absolutely open design protects your investment with maximum flexibility for future extensions. It hosts the experimental setup in a more spacious enclosure that also provides room for rotating anode, revolutionary liquid metal X-ray source for structural biology, highest X-ray intensity in-house for even smaller and more challenging samples, self-regenerating target with Ga-radiation for lower background scattering and radiation damage, combination of two different wavelengths, fast and fully-automated wavelength switch.
The D8 VENTURE hosts the experimental setup in a more spacious enclosure that also provides room for rotating anode, liquid metal jet and dual wavelengths solution. All systems feature the revolutionary PHOTON II CPAD detector.
The D8 VENTURE offers highest experimental flexibility, with excellent sample accessibility and visibility. It is engineered with high modularity from best-in-class components and is available in popular all-air-cooled configurations. The absolutely open design protects your investment with maximum flexibility for future extensions.
Configuration: θ-θ horizontal Bragg-Brentanno geometry
Goniometer: 600mm circle goniometer
Source: CuKa (1.6kW max)
Primary optics: fixed slits, knife edge collimator
Diffracted optics: fixed antiscatter slits, Cu absorber, soller slits, monochromator
Detectors: Scintillation, 1-D solid state super fast detector (Lynx-Eye)
Parallel beam optics: Gobel mirror
Available options: Grazing incidence XRD, Thin film measurements (XRR), high temperature furnace (RT-900oC) working under reductive and oxidative atmospheres
Professor Elias Eliopoulos is the Director of the Genetics Laboratory at the Biotechnology Department of the Agricultural University of Athens (A.U.A.) and leads the bioinformatics and structural biology group. He is professor of Biochemistry, a biophysicist and crystallographer by training with considerable experience in biomolecular structure analysis, epitope mapping of protein receptors, genetic polymorphisms and variation, cloning, with experimental and computational methods, protein structure prediction, ligand and drug design, protein design, in silico antibody design and biosoftware development. The group has experience and international reputation on computational protein folding, ab initio and homology modeling of proteins derived from edge gene research and membrane protein modeling. The group is also involved in many ligand computational design projects for pharmaceutical, medicinal and agricultural applications.
Server consists of 24 units of 8-core Intel Xeon 2.8GHz workstations with 32GB internal memory each.
The D8 Venture system includes:
IμS microfocus X-ray source, Cu radiation, air cooled
Four-circle goniometer, computer-controlled, crystal-detector distance = 35-240mm, sphere of confusion of less than 7 micrometer
Detector PHOTON 100, CMOS technology, air-cooled, 100 x 100mm active area, 240e/Cu sensitivity, shutterless data collection
800 Series Cryostream cooler, Oxford Cryosystems, AD51 Dry Air Unit, Integrated Autofill system, 60 lt and 150 lt dewar vessels.
APEX3 and PROTEUM software
EDUCATION
2019: PhD in Structural Biology and Protein Biomaterials (UoC, IMBB-FORTH - Greece)
2012: M.Sc. in Protein Biotechnology (UoC-Greece)
2009: Diploma Degree in Biology (UoC-Greece)
WORKING EXPERIENCE
Oct 2019 – Now: Post-Doctoral Researcher at I.M.B.B.-F.O.R.TH, Greece
(Laboratory of Structural Biology)
Sep 2020 – Now: Autonomous teaching and exam organizing of the under-graduate courses
"Enzymatic Biotechnology" and “Protein Engineering” at the Dept. of
Biology of the Uni. of Crete
2014 – 2020: Experienced scientist position at I.M.B.B.-F.O.R.TH, Greece (InnovCrete project /FP7-Regpot program) and at University of Crete
Oct. 2010 –Jan. 2011: Laboratory Teaching Assistant of Microbiology Teaching Lab
at the Dept. of Biology, Uni. of Crete
SELECTED PUBLICATIONS
*Kefala K., Kotsifaki D., Providaki M., Kapetaniou E., Rahme L. and Kokkinidis M. Purification, crystallization and preliminary X-ray diffraction analysis of the C-terminal fragment of the MvfR protein of Pseudomonas aeruginosa. (2012). Acta. Cryst F, 68, 695-697
*Kefala A., Kotsifaki D., Providaki M., Amprazi M. and Kokkinidis M. Expression, purification and crystallization of a protein resulting from the inversion of the aminoacid sequence of a helical bundle. (2017). ). Acta. Cryst F, 73, 51-53
Dimovasili C., Fadouloglou V.E., Kefala A., Providaki M., Kotsifaki D, Sarrou I., Plaitakis A., Zaganas I. and Kokkinidis M. Crystal structure of glutamate dehydrogenase 2, a positively selected novel human enzyme involved in brain biology and cancer pathophysiology. Journal of Neurochemistry (accepted)
RESEARCH PROFILE
Being in a Structural Biology group since 2011, Dr. Kefala A. has studied deeply the protein folding problem, elucidating the relation between amino acid sequence and structural properties of α-helical bundles. The knowledge obtained was used for protein design leading to bio-derived materials for bio-medical applications and basic research through structural studies of protein folding through reverse sequences.
Notable achievements include:
The elucidation of the protein folding of mutants of the α-helical protein Rop that is used as a canonical paradigm of this structural motif.
Protein design of novel protein-based biomaterials with specific engineered properties.
The D8 VENTURE hosts the experimental setup in a more spacious enclosure that also provides room for rotating anode, liquid metal jet and dual wavelengths solution. All systems feature the revolutionary PHOTON II CPAD detector.
The D8 VENTURE offers highest experimental flexibility, with excellent sample accessibility and visibility. It is engineered with high modularity from best-in-class components and is available in popular all-air-cooled configurations. The absolutely open design protects your investment with maximum flexibility for future extensions.