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Using the SIFT tool, the result was "Damaging" to protein function [ 11].
In silico analysis of the W210R mutation using the SIFT tool (http://sift.jcvi.org; PMID: 19561590) and the PolyPhen-2 tool (http://genetics.bwh.harvard.edu/pph2/index.shtml; PMID: 20354512) consistently showed a probably damaging effect on the protein, reaching highest possible scores of the two algorithms (SIFT score: 0.00; PolyPhen-2 score: 1.000).
Notably, all of the amino acid mutations produced by the missense mutations are predicted to be mutagenic, using the SIFT tool, which sorts intolerant from tolerant substitutions on the basis of conservation of the particular amino acid in related proteins (Ng and Henikoff 2001).
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Variant functional impacts were predicted using the SiFT Blink tool (http://sift.jcvi.org/www/SIFT_BLink_submit.html) (Kumar et al. 2009).
A possible effect of the validated SNPs was analyzed using the SIFT and CFSSP prediction tools.
The deleterious effect of non-synonymous SNPs were analysed using the SIFT and PolyPhen algorithms.
SIFT: The follistatin protein sequences were submitted as input for the SIFT tool.
SIFT (Sorting Intolerant from Tolerant): The follistatin protein sequences were submitted as input for the SIFT tool.
Then, we use the SIFT extractor to extract the SIFT features from all training spectrogram images.
The effect of the p. (Asp92Gly) substitution on SDHD function was predicted using the in silico tools SIFT (Ng and Henikoff 2003), Align GVGD (Tavtigian et al. 2006) and Polyphen (Adzhubei et al. 2010), all running recommended parameters.
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