Member Site › Forums › Rosetta 3 › Rosetta 3 – General › Comparatively modelled chains are far apart in final structure while the template pdb has all chains in a close-knit complex
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November 1, 2020 at 7:00 pm #3615Anonymous
I tried to model a protein sequence (4 chains) using RosettaCM. The structure of the protein is already available in PDB (2ERJ). The issue I am facing – the final modelled structure has all the 4 chains very far apart, unlike the pdb. What exactly is going wrong?
I am obtaining the alignment file from Clustal omega. The target fasta sequence is given as chain1/chain2/chain3/chain4 and the 2ERJ fasta is also given in the same format in clustal omega so that the final alignment has only 2 unique identifiers in the ali file, like this:
2ERJ1|Chains GMLSLELCDDDPPEIPHATFKAMAYKEGTMLNCECKRGFRRIKSGSLYMLCTGSSSHSSW 60
IL2Ra
ELCDDDPPEIPHATFKAMAYKEGTMLNCECKRGFRRIKSGSLYMLCTGNSSHSSW 55************************************************.******
2ERJ1|Chains DNQCQCTSSATRSTTKQVTPQPEEQKERKTTEMQSPMQPVDQASLPGHCREPPPWENEAT 120
IL2Ra DNQCQCTSSATRNTTKQVTPQPEEQKERKTTEMQSPMQPVDQASLPGHCREPPPWENEAT 115
************.***********************************************
2ERJ1|Chains ERIYHFVVGQMVYYQCVQGYRALHRGPAESVCKMTHGKTRWTQPQLICTGEMETSQFPGE 180
IL2Ra ERIYHFVVGQMVYYQCVQGYRALHRGPAESVCKMTHGKTRWTQPQLICTGEMETSQFPGE 175
************************************************************
2ERJ1|Chains EKPQASPEGRPESETSCLVTTTDFQIQTEMAATMETSTGHHHHH
224IL2Ra EKPQASPEGRPESETSCLVTTTDFQIQTEMAATMETSIFTTEYQVAVAGCVFLLISVLLL 235
************************************* .::
and so on.
I used the following commands:
~wholepath/tools/protein_tools/scripts/setup_RosettaCM.py –fasta IL2Ra.fasta –alignment alignmentfile.aln –alignment_format clustalw –templates 2ERJ.pdb
~wholepath/main/source/bin/rosetta_scripts.default.linuxgccrelease @flags -database /wholepath/main/database -nstruct 10
What is it that I am doing wrong which is leading to all the modelled chains being very far apart in the final structures?
Kindly help.
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