Structural & Supramolecular Chemistry Research Group – Research – New crystallisation methodology for biological macromolecules

D. New crystallisation methodology for biological macromolecules

Research into substances and materials promoting the heterogeneous nucleation of macromolecular crystals is a blossoming topic in crystallogenesis. We have participated in developing the use of materials containing pores or cavities, such as Bioglass, carbon nanotubes, and Molecularly Imprinted Polymers (MIPs), as heterogeneous nucleants in collaboration with the Group of Prof. Naomi Chayen, Imperial College London and are pursuing further ideas.D1-

Figure 1. Human macrophage Migration Inhibiting Factor grown in the presence of MIPs

Research on other techniques for effective a priori prediction of crystallisation conditions is also being actively pursued: (a) As coordinators of the Industry-Academia Partnerships and Pathways – Marie Curie Project TOPCRYST, we developed the use of Dual Polarization Interferometry, pioneered by Farfield Scientific Ltd., to probe crystallisation at its most crucial stages. This allows to predict the outcome of crystallisation trials when they are still at their earliest stages and thus to rationally design and direct such experiments in order to lead them to well-diffracting crystals.D7 (b) Moreover, the use of Genetic AlgorithmsD8 and of calorimetryD9 are additional techniques that have been tested.

  1. Protein crystallization: from purified protein to diffraction-quality N. E. Chayen, E. Saridakis, Nat. Methods 2008, 5, 147-153.
  2. Carbon-nanotube based materials for protein crystallization. P. Asanithi, E. Saridakis, L. Govada, I. Jurewicz, E.W. Brunner, R. Ponnusamy, J.A.S. Cleaver, B. Dalton, N.E. Chayen,  R.P. Sear, ACS Appl. Mat. Interf. 2009, 1, 1203-1210.
  3. Protein crystallization facilitated by molecularly imprinted polymers. E. Saridakis, S. Khurshid, L. Govada, Q. Phan, D. Hawkins, G.V. Crichlow, E. Lolis, S.M. Reddy and N.E. Chayen, Natl. Acad. Sci. USA 2011, 108, 11081-11086.
  4. Imprinted polymers assisting protein crystallization. Saridakis, N. E. Chayen Trends Biotech. 2013, 31, 515-520.
  5. Khurshid, E. Saridakis, L. Govada and N.E. Chayen. “Porous nucleating agents for protein crystallization.” Nat. Protoc. (2014) 9, 1621-1633.
  6. Protein crystal nucleation in pores. N. Nanev, E. Saridakis, N. E. Chayen, Scientific Reports 2017, 7: 35821.
  7. Use of Dual Polarisation Interferometry as a diagnostic tool for protein crystallization. Boudjemline, E. Saridakis, M. J. Swann, L. Govada, I. M. Mavridis, N. E. Chayen, Anal. Chem. 2011, 83, 7881-7887.
  8. A novel Genetic Algorithm-inspired concept for macromolecular crystal optimisation. E. Saridakis. Crystal Growth Des. 2011, 11, 2993-2998.
  9. Influence of precipitating agents on thermodynamic parameters of protein crystallization solutions. P. Stavros, E. Saridakis, G. Nounesis Biopolymers 2016, 105, 642-652.