Modeling a Homodimer using a Monomer Created in HM (video)
This is a basic method for any oligomer. More complex protocols with more automation may also be introduced, but this is a generally applicable method that allows you flexibility, assuming that when you look at step (I.1) the homologs do not extensively interpenetrate but are separate globular proteins.
NOTE: This workflow will also work for multi-chain Homology Modeling. For step II, find a template where there’s a homologous structure of something else binding to your protein of interest. Once you find this template, continue with this workflow, using this template in the place of 2Q8S.
Example for a C2 Symmetric Homodimer (Hamster PPAR)
Predict Monomer Structure
- Paste target monomer sequence (e.g. Hamster PPAR) in Cyrus HM and run Homology Modeling
- Download PDB of top resulting model (Monomer Model)
Find Homodimer Template
- Run blastp on target sequence (e.g. HamsterPPAR) against the PDB
- Based on your specific knowledge and literature, select the most relevant homodimer PDB structure as a template for dimerization (e.g. 2Q8S human PPAR complex)
- Download 2Q8S PDB and reserve for later
- Method requires a template with your desired symmetry
Pre-Symmetrize Model
- In PyMOL, superimpose Monomer Model to Chain A of template 2Q8S and an identical copy of Monomer Model to Chain B of template 2Q8S
- Save each of the aligned Monomer Models from PyMOL to two separate PDBs
- Using a text editor, combine the two PDBs with a TER line in between, and by changing Chain identifier from A to B in one of the monomers. This is the Pre-Symmetrized Model
Optimize Side Chains
- Load the Pre-Symmetrized Model into Cyrus CAD. This needs side chains fixed
- Run Relax or Repack with five or more repetitions to optimize side chains. This places side chains realistically.
- Select structure with lowest energy. This is your final Homodimer Model!
While this method is used for a C2 symmetric homodimer in the example above, it can also be used to template oligomeric models off of any template regardless of symmetry.