Applications of Homology Modeling Hanka Venselaar.
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Transcript of Applications of Homology Modeling Hanka Venselaar.
Homology modeling
Example: by 80 residues 30% identity sufficient
# residues
% id
entit
y
*
* Actually, modelling is possible, but we cannot get an alignment…
O
Homology modeling in short…Prediction of structure based upon a highly similar structure
Add sidechains, Molecular Dynamics simulation on model
Unknown structure
NSDSECPLSHDG
NSDSECPLSHDG
|| || | ||
NSYPGCPSSYDG
Alignment of model and template sequence
Known structure
Known structure Back bone copiedCopy backbone and conserved residues
Model!
1: Template recognition and initial alignment
• BLAST your sequence against PDB
• Best hit normally template
• Initial alignment
NSDSECPLSHDGYCLHDGVC
|| || | ||||| |||
NSYPGCPSSYDGYCLNGGVC
2: Alignment correction
• Functional residues conserved• Use multiple sequence alignments• Deletions shift gaps
CPISRTGASIFRCW CPISRTGASIFRCWCPISRTA---FRCW CPISRT---AFRCW
CPISRTAAS-FRCWCPISRTG-SMFRCWCPISRTA--TFRCWCPISRTAASHFRCWCPISRTGASIFRCW CPISRTA---FRCW
Both are possible
Multipe sequence alignment
Correct alignment
Sequence with known structure
Your sequence
2: Alignment correction
• Core residues conserved• Use multiple sequence alignments• Deletions in your sequence shift gaps
Known structure FDICRLPGSAEAV
Model FNVCRMP---EAI
Model FNVCR---MPEAI
S
G
P
L
A
E
R
C
I V
C
R
M
P
EV
C
R M
P
E
Correct alignment
F-D--A-V
3: Backbone generation
• Making the model….• Copy backbone of template to model• Make deletions as discussed• (Keep conserved residues)
1: Template recognition and initial alignment
2: Alignment correction
3: Backbone generation
4: Loop modeling
4: Loop modeling
Known structure GVCMYIEA---LDKYACNC
Your sequence GECFMVKDLSNPSRYLCKC
Loop library,
try different options
1: Template recognition and initial alignment
2: Alignment correction
3: Backbone generation
4: Loop modeling
5: Sidechain modeling
5: Side-chain modeling
• Several options
• Libraries of preferred rotamers based upon backbone conformation
1: Template recognition and initial alignment
2: Alignment correction
3: Backbone generation
4: Loop modeling
5: Sidechain modeling
6: Model optimization
6: Model optimization
• Molecular dynamics simulation• Remove big errors
• Structure moves to lowest energy conformation
1: Template recognition and initial alignment
2: Alignment correction
3: Backbone generation
4: Loop modeling
5: Sidechain modeling
6: Model optimization
7: Model validation
7: Model Validation
• Second opinion by PDBreport /WHATIF
• Errors in active site? new alignment/ template
• No errors? Model!
1: Template recognition and initial alignment
2: Alignment correction
3: Backbone generation
4: Loop modeling
5: Sidechain modeling
6: Model optimization
7: Model validation
8: Iteration
8: Iteration
8: Iteration
8: Iteration
Model!
1: Template recognition and initial alignment
2: Alignment correction
3: Backbone generation
4: Loop modeling
5: Sidechain modeling
6: Model optimization
7: Model validation
8: Iteration
8: Iteration
8: Iteration
8: Iteration
8 steps of homology modeling
1: Template recognition and initial alignment2: Alignment correction3: Backbone generation4: Loop modeling5: Side-chain modeling6: Model optimization7: Model validation8: Iteration
Alignment
Modeling
Correction
Homology Modeling…• What? Prediction of an unknown structure based on
an homologous and known structure• Why? To answer biological and medical questions
when the “real” structure is unknown• When? A template with enough identity must be
available• How? 8 Steps
To conclude….