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Team:EPF-Lausanne/Modeling overview
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So the aim of the molecular dynamics simulation is to simulate the LOV domain in its environment under light activation (so-called light state) and without light activation (ground state, so-called dark state), calculate atom and residue movements of particular/interesting LOV domain regions, and finally deduce which residue(s) could be mutated to stabilize the light-activated state of this LOV domain (increase its lifetime). | So the aim of the molecular dynamics simulation is to simulate the LOV domain in its environment under light activation (so-called light state) and without light activation (ground state, so-called dark state), calculate atom and residue movements of particular/interesting LOV domain regions, and finally deduce which residue(s) could be mutated to stabilize the light-activated state of this LOV domain (increase its lifetime). | ||
| - | Then, simulation of the complete LOVTAP protein with selected mutations could give us insights about the | + | Then, simulation of the complete LOVTAP protein with selected mutations could give us insights about the behaviour of our protein in its environment. |
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Revision as of 13:20, 18 October 2009
Contents |
Protein domain of interest
Our protein of interest is LOVTAP. This protein was synthetically engineered by Pr. [http://www.ncbi.nlm.nih.gov/sites/entrez?db=pubmed&cmd=search&term=18667691 Sosnick's] group from the University of Chicago. It is a fusion protein between a LOV domain (Avena Sativa phototropin 1) and the E. Coli tryptophan repressor. This protein undergoes changes under light activation as shown by Sosnick et al, namely when the protein is activated by light it binds to DNA and inversely.
For more information about LOVTAP protein please click here.
Goal
Sosnick et al. found that light-activated LOVTAP isn't stable. After light excitation, the LOV domain returns to its ground state (non light-activated state) very quickly.
So the aim of the molecular dynamics simulation is to simulate the LOV domain in its environment under light activation (so-called light state) and without light activation (ground state, so-called dark state), calculate atom and residue movements of particular/interesting LOV domain regions, and finally deduce which residue(s) could be mutated to stabilize the light-activated state of this LOV domain (increase its lifetime).
Then, simulation of the complete LOVTAP protein with selected mutations could give us insights about the behaviour of our protein in its environment.
Starting material
Both LOV domain crystallography files were obtained from [http://www.rcsb.org/pdb/home/home.do RCSB]:
- [http://www.rcsb.org/pdb/explore/explore.do?structureId=2V0W Light-activated LOV domain]
- [http://www.rcsb.org/pdb/explore/explore.do?structureId=2V0U Dark LOV domain]
These crystallographies were done by [http://www.ncbi.nlm.nih.gov/pubmed/18001137 Halavaty et al.].
Molecular dynamics: a little theory
Molecular dynamics simulation consists in the numerical, step-by-step, solution of the classical equations of motion. For this purpose we need to be able to calculate the forces acting on the atoms, and these are usually derived from a potential energy.
StepsThe following information is mostly taken from an Introduction to Molecular Dynamics: see [http://chsfpc5.chem.ncsu.edu/~franzen/CH795N/lecture/IV/IV.html here] their web page. 1. MinimizationUsing the forcefield that has been assigned to the atoms in the system, it is essential to find a stable point or a minimum on the potential energy surface in order to begin dynamics. At a minimum on the potential energy surface, the net force on each atom vanishes.
2. EquilibrationMolecular dynamics solves the equations of motion for a system of atoms. The solution for the equations of motion of a molecule represents the time evolution of the molecular motions, the trajectory. Depending on the temperature at which a simulation is run, molecular dynamics allows barrier crossing and exploration of multiple configurations.
3. Analysis and validationThis part is dedicated to the analysis of our previous results, in order to validate the following researches. For more details about what we have done, see :
4. SimulationWe run a 100ns simulation, from which we will collect the data and see what happens to our protein! We made calculations during nearly 4 weeks, on 64 processors.
5. Atom movement analysisIn this last section we analyze the atom movement using the PCA analysis (Principal Component Analysis), for making predictive models.
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