Albertine Donker

IRIDA: a Heterogeneous Disease 181 4 Supplemental Table 5. Continued a http://exac.broadinstitute.org/variant/22-37492064-CA-C b Align GVGD, web based in silico prediction software program that combines the biophysical characteristics of amino acids and protein multiple sequence alignments to predict where missense substitutions in genes of interest fall in a spectrum from enriched delterious to enriched neutral. A-GVGD scores amino acid substitutions on a 7-scale scoring system, from C0 to C65. An amino acid substitution with a C0 score is considered to be neutral, amino acids with C15 and C25 scores are considered intermediate, as changes to protein structure or function are uncertain, and C35 scores or higher are considered as likely deleterious. 9 c PolyPhen-2 (Polymorphism Phenotyping v2 HumVar) is a tool, which predicts possible impact of an amino acid substitution on the structure and function of a human protein using straightforward physical and comparative considerations. PolyPhen scores range from 0 ≤ 1 ≤ X , outcome scores of 0.00–0.15 are classified as benign, 0.15–1.0 as possibly damaging, 0.85-1.0 as more confidently predicted to be damaging. 10 d The SIFT algorithm combines sequence homology and physical properties of amino acid substitutions to analyze whether or not amino acid substitutions are tolerated, in light of the predicted effect on the protein structure. SIFT score ranges from 0 to 1. The amino acid substitution is predicted damaging if the score is < 0.05 and tolerated if the score is > 0.5 11 e http://exac.broadinstitute.org/variant/22-37471208-G-A f http://exac.broadinstitute.org/variant/22-37465149-A-G g http://exac.broadinstitute.org/variant/22-37462173-C-T h This substitution is predicted to introduce a new and more efficient acceptor splice site 4 bases downstream from the original acceptor splice site in intron 2 leading to a frameshift in the open reading frame. Not proven on RNA or protein level. (http://www.interactive-biosoftware.com/doc/alamut- visual/2.7/splicing.html), accessed on July 14 2016. i This substitution is located in the donor splice site of intron 7. The consequence of this change is not predictable, but a skip of exon 7 is very likely resulting in the loss of a classical splice site. Not proven on mRNA or protein level (http://www.interactive-biosoftware.com/doc/alamut-visual/2.7/splicing.html) , accessed on July 14 2016. Abbreviations: na denotes not applicable