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H2020Doctoral network2017–2021

RAMP · RAtionalising Membrane Protein crystallisation

Horizon 2020 — Marie Skłodowska-Curie Actions

Duration
2017-03-01 → 2021-08-31
EU contribution
€3,240,526
Participants
10
Scheme
MSCA-ITN-ETN

Lines connect the coordinator with its partners.

Results in brief

RAtionalising Membrane Protein crystallisation

Over the past two decades, advances in the crystallisation of soluble proteins, diffraction data collection and data analysis have made structure solution of soluble proteins almost routine. But crystallisation – remains a major bottleneck for membrane proteins. The additional challenge with membrane proteins is two-fold: first, the protein itself is hard to produce in large quantities and tends to be unstable. Second, the parameter space is even larger than for soluble proteins due to additional parameters: the detergents. The crystallisation process is thus complex and poorly understood. Membrane proteins are currently crystallized by using brute-force screening to search this high-dimensional parameter space to find initial conditions, followed by trial-and-error optimisation to grow crystals suitable for diffraction studies. Though over 85% of drug targets are membrane proteins, fewer than 650 unique membrane protein structures have been determined. We urgently require better methods to crystallize membrane proteins. RAMP created a unique training network that brings together three strands: (1) development of a microfluidics-based technology to control membrane protein crystallisation, and the ability to sample the large parameter space of crystallisation conditions rapidly; (2) the introduction to membrane protein crystallisation optimisation of modelling of the phase diagram, a technique used with great success for soluble protein crystallisation; and (3) the application of these developments to medically and biologically important membrane protein targets. RAMP also trained the students on two new and emerging techniques: 1) serial crystallographic methods. These are increasingly used at synchrotron sources as well as at rapidly developing ultra-bright free-electron laser sources, and require crystals in the 1-20 μm size to solve structure of previously intractable proteins. 2) Neutron protein crystallography, which requires large crystals (> 0.01 mm3). It is, however, the only way to visualise protons - important information for drug design.

Data: CORDIS, © European Union

Project objective

Membrane proteins form more than 85% of drug targets, but just 600 unique membrane protein crystal structures have been determined. A better understanding of how to crystallize membrane proteins reliably is therefore urgently required. The Innovative Training Network “RAtionalising Membrane Protein crystallisation” – RAMP will bring together cutting-edge physical chemistry methods for crystallisation condition control and phase diagram exploration, and the development of new lipids and screens in conjunction with industry with the most challenging biological problems. The network includes expert academic and industrial research groups in crystallisation theory, methods development, membrane protein crystallography, drug development and novel structural techniques like time-resolved and neutron crystallography. We will develop new, rational methods for crystallising membrane proteins, focusing particularly on transporters that are also interesting drug targets. The new robust crystallisation methods will also allow us to use emerging European research infrastructures like XFEL or ESS to gain insight into membrane protein function because the techniques will provide the necessary precise control of crystal size. A structured training programme organized by academia and industry together will equip the early stage researchers with the skills needed for a successful research career in the field of structural biology. Frequent secondments, research visits and meetings between early-career scientists ensure an efficient exchange of ideas and practical experiences between different groups leading to better integration of European research and innovations in structural biology. Supervision and mentoring by several senior scientists will give the researchers a strong scientific education and make them highly competitive in the work place of tomorrow. The work programme here will improve European competitiveness and advance graduate training.

Original text from CORDIS.

Participants

  • UNIVERSITE GRENOBLE ALPES · GrenobleCoordinatorFrance
  • AARHUS UNIVERSITET · Aarhus CDenmark
  • EUROPEAN SPALLATION SOURCE ERIC · LundSweden
  • IMPERIAL COLLEGE OF SCIENCE TECHNOLOGY AND MEDICINE · LondonUnited Kingdom
  • NATIONAL UNIVERSITY OF IRELAND MAYNOOTH · MaynoothIreland
  • THE PROVOST, FELLOWS, FOUNDATION SCHOLARS & THE OTHER MEMBERS OF BOARD, OF THE COLLEGE OF THE HOLY & UNDIVIDED TRINITY OF QUEEN ELIZABETH NEAR DUBLIN · DublinIreland
  • UNIVERSITY OF BRISTOL · BRISTOLUnited Kingdom
  • UNIVERSITY OF HAMBURG · HamburgGermany
  • UNIVERSITY OF LEEDS · LeedsUnited Kingdom
  • UNIVERSITY OF SURREY · GuildfordUnited Kingdom

Links

Data: CORDIS, © European Union