Supporting Information
to
Eight novel F13A1 gene missense mutations in patients with mild FXIII deficiency: In sillico analysis suggests pathologic changes in FXIII A-subunit structure/function
Biswas A1*§, Ivaskevicius V1*§, Thomas A1, Varvenne M2, Brand B3, Rott H4, Haussels I1,
Ruehl H1, Oldenburg J1
1 Institute of Experimental Haematology and Transfusion Medicine, University Clinic Bonn, Bonn, Germany
2 Department of Hematology, Hemostasis, Oncology and Stem Cell Transplantation, Hannover Medical School, Hannover, Germany
3 Division of Haematology, University Clinic Zurich, Zurich, Switzerland
4 Coagulation CenterRhein-Ruhr, Duisburg, Germany
*AB and VIcontributed equally to this manuscript.
The online version of this article contains a supplementary appendix.
§Correspondence:
VytautasIvaskevicius, Institute of Experimental Haematology and Transfusion Medicine, University Clinic Bonn, Sigmund-Freud-Str. 25, 53127 Bonn, Germany,
e-mail:
phone.: +49-228-287-15188
fax.: +49-228-287-14320
or
Arijit Biswas, Institute of Experimental Haematology and Transfusion Medicine,
University Clinic Bonn, Sigmund Freud Str. 25, 53127 Bonn, Germany.
Email:
phone: +49 228 287 19428
fax: +49 228 287 14320
Figure S1. Structural alignment of activated and zymogenic FXIII structure.
Footnotes to Fig S1:Panel A shows the structural alignment graph for the PDB files 4kty and 1f13 created on the FATCAT structural alignment server. The major twists between the two structures are reperesented by arrows in this image. Panel B shows the structurally aligned monomers of zymogenic FXIII (Cyan) and activated FXIII (yellow) depicted in ribbon format (PDB files 1f13 and 4kty respectively). Structural alignment was generated on the TM-align server.
Figure S2. Conservation of the His342 (His342Tyr) residue.
Footnotes to Fig S2: The image shows the multiple alignment of FXIII A subunit with FXIII A from other species and also with other known Transglutaminases. The region of FXIII A shown includes the His342 residue which is highly conserved across both types of alignment.
Figure S3.Aggregation profile for the FXIII A subunit amino acid sequence
Footnotes to Fig S3: The graph shows regions of FXIII A subunit which are more prone to aggregation. The peaks represent highly aggregation prone regions.
Figure S4.Disordered regions in FXIII A subunit amino acid sequence
Footnotes to Fig S4: The image shows the prediction of disordered regions in FXIII A subunit. The peaks represent the degree of disorder. The red line indicates the minimum threshold for identification of a disordered region with a false positive prediction rate of 5%.
Table S1. Atomic details of Hydrogen bonds for the respective residues.
Footnotes to Table S1: The Table lists all the H-bonds/salt bridges (observed in the post simulation averagedzymogenic structure) formed by the residues on which the mutations have been reported in this study.
Table S2. Pathogenicity prediction results for the F13A1 mutations.
Mutation / SNPs&GO / Polyphen-2 / MutPredPro166Leu / Neutral / Damaging / Non-Deleterious
Arg171Gln / Disease / Damaging / Non-Deleterious
His342Tyr / Disease / Damaging / Deleterious
Gln415Arg / Neutral / Benign / Non-Deleterious
Leu529Pro / Disease / Damaging / Deleterious
Gln601Lys / Disease / Damaging / Deleterious
Arg703Gln / Disease / Damaging / Deleterious
Arg715Gly / Neutral / Damaging / Non-Deleterious
Footnotes to Table S2: Three pathogenicity predictions servers were queried for the reported mutations [ on 05.02.2013, on 05.02.2013, on 05.02.2013].
Table S3. Prediction of Ubiquitination sites on the mutated FXIII A amino acid sequence.
Footnotes to Table S3: This table lists the predicted Ubiquitination sites in the FXIII A subunit amino acid sequence with their likely possibilities represented as a scoring function. The threshold for this scoring function all across is -1.90.
1