Fabry:Homology based structure predictions/Journal

Fabry Disease » Homology based structure predictions » Journal

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Dataset preparation

The homology search was performed online and resulted in the two output files hhpred.out and coma.out for the structure search with HHpred and COMA, respectively. In both cases, we used the default values, thresholds and databases. From both the resulting files we tried in each case to create three distinct datasets with the demanded sequence identity to the target protein with the following calls and scripts.

perl make_dataset_hhpred.pl hhpred.out 0.0000000000000001

This resulted in the HHpred datasets mentioned in Dataset preparation Table 1 and the corresponding pdb structure files.

perl make_dataset_coma.pl coma.out 0.002

This resulted in the Coma datasets mentioned in Dataset preparation Table 2 and the corresponding pdb structure files.

Calculation of models

Modeller

Default settings

For the standard homology modeling with Modeller two basic scripts were used, which build up on the ones described in the recommended tutorial: 1_align.py and 2_Single_template_modeling.py (in this case example file for 1ktb) which need the appropriate template pdb structure files, as well as the target (AGAL) sequence in pir format as input:

>P1;1R46
sequence:1R46:::::::0.00: 0.00
MQLRNPELHLGCALALRFLALVSWDIPGARALDNGLARTPTMGWLHWERFMCNLDCQEEP
DSCISEKLFMEMAELMVSEGWKDAGYEYLCIDDCWMAPQRDSEGRLQADPQRFPHGIRQL
ANYVHSKGLKLGIYADVGNKTCAGFPGSFGYYDIDAQTFADWGVDLLKFDGCYCDSLENL
ADGYKHMSLALNRTGRSIVYSCEWPLYMWPFQKPNYTEIRQYCNHWRNFADIDDSWKSIK
SILDWTSFNQERIVDVAGPGGWNDPDMLVIGNFGLSWNQQVTQMALWAIMAAPLFMSNDL
RHISPQAKALLQDKDVIAINQDPLGKQGYQLRQGDNFEVWERPLSGLAWAVAMINRQEIG
GPRSYTIAVASLGKGVACNPACFITQLLPVKRKLGFYEWTSRLRSHINPTGTVLLQLENT
MQMSLKDLL*

The following runs were performed on our home computers:

#3HG3 100%
mod9.10 1_align_3hg3.py
mod9.10 2_Single_template_modeling_3hg3.py

#1KTB 53%
mod9.10 1_align.py
mod9.10 2_Single_template_modeling.py

#3CC1 25%
mod9.10 1_align_3cc1.py
mod9.10 2_Single_template_modeling_3cc1.py

Multiple templates

For multiple templates three scripts are needed, which again base on the tutorial and are here again only exemplified for one case: 3_multiAli_1.py, 4_add1R46_1.py and 5_multipleMod_1.py The following combinations were performed:

#Multiple 3HG3,1KTB
mod9.10 3_multiAli_1.py
mod9.10 4_add1R46_1.py
mod9.10 5_multipleMod_1.py

#Multiple 3HG3,1KTB,3CC1
mod9.10 3_multiAli_2.py
mod9.10 4_add1R46_2.py
mod9.10 5_multipleMod_2.py

#Multiple 3CC1, 3ZSS, 3A24
mod9.10 3_multiAli_3.py
mod9.10 4_add1R46_3.py
mod9.10 5_multipleMod_3.py

#Multiple 3CC1, 3HG3
mod9.10 3_multiAli_4.py
mod9.10 4_add1R46_4.py
mod9.10 5_multipleMod_4.py

Edited Alignment input

For the edited alignment models, only the second Modeller script 2_Single_template_modeling.py was needed. Edited alignments were provided as input. We rearranged them with the program SeaView.

#Active Site shifted right to next D (7 and 1) in -2d.ali
mod9.10 2_Single_template_modeling_3hg3_changed_actSite.py

#Active Site shifted right to next D (7 and 1) in both ali files
mod9.10 2_Single_template_modeling_3hg3_changed_actSite_2.py

#Active Site shifted right to next D (7 and 1) in both ali files + Substrate binding region (203-207) forced to be consecutive
mod9.10 2_Single_template_modeling_3hg3_changed_actSite_3.py