Table 6.

In silico analysis of blood Factor VIII (FVIII) (PDB ID: http://www.rcsb.org/pdb/search/structidSearch.do?structureId=2R7E, UniProt ID: http://www.uniprot.org/uniprot/P00451; GenBank: NM_000132.3)

FVIII Amino acid/mutation	DUET ΔΔG, kcal/mol	PopMusica ΔΔG, kcal/mol	SAAFEC ΔΔG, kcal/mol	MAESTROwebb ΔΔG, kcal/mol	PolyPhen‐2 Score/mutation prediction	MSAc aa conservation	Involved in predicted ligand‐binding pockets/pocket No. (FTMap)	Surface exposure (%)d	Involved in predicted protein–protein interaction sites (meta‐PPISP)	Node degree (RING‐2.0)	Predicted fluctuation value (FlexPred)	Flexibility classe (PredyFlexy)	Experimental data	References
Ala2201 A2201P	0.378 Stabilizing	0.23 Destabilizing SA = 13.5%	−0.985608 Destabilizing	0.196 Destabilizing Cpred = 0.887	0.988 Probably damaging	High	No	36	Yes	4	0.075	1	Mutation associated with mild hemophilia A The mutant protein domain in vitro seems to have molecular functions similar to the wild type (investigated properties: stability, and binding to the vWF) but membrane binding is damaged	Pratt et al. (1999); Liu et al. (2000). Spiegel et al. (2004)
Met2238 M2238V	−0.503 Destabilizing	0.27 Destabilizing SA = 10.0%	0.553310 Increase Stability	0.248 Destabilizing Cpred = 0.874	0.152 Benign	High	No	11	No but very close to a proposed site	8	1.487	1	Mutation associated with moderate hemophilia A The mutant protein domain in vitro seems to have molecular functions similar to the wild type (investigated properties: stability, membrane binding and binding to the vWF) (in vitro)	Pratt et al. (1999); Liu et al. (2000). Spiegel et al. (2004)

^aFor the program PopMusic solvent accessibility (SA) values are shown (in percent).

^bFor the program MaestroWeb the confidence estimation Cpred is shown (0.0‐not reliable and 1.0‐highly reliable).

^cMSA‐Multiple sequence alignment.

^dLiu et al. (2000).

^eFlexibility class was determined by the program PredyFlexy (rigid‐0, intermediate‐1, flexible‐2).

Wild‐type residue (bold) and amino acid substitution (Underlined).