Table 1. VUSs enriched in patients with an abnormal NBS suggestive of VLCADD.

VUS	Allele count*	Expected count¶	P-value¥	Previously reported	SIFT / Polyphen-2$	Structural location§	Summary of results
c.1066A>G (p.I356V)	11	1.3-13.9	2.38×10-7 to 0.56	no	D/D	alpha-helical bundle	unclear
c.1273G>A (p.A425T)	8	0.05-0.5	1.24×10-13 to 1.16×10-5	(Chien et al 2013)	T/D	alpha-helical bundle	likely pathogenic
c.1001T>G (p.M334R)	6	0.03-0.06	1.71×10-10 to 5.75×10-9	no	D/D	near active site	likely pathogenic
c.538G>A (p.A180T)	5	0.02-0.12	3.79×10-9 to 7.65×10-5	no	D/D	near active site	likely pathogenic
c.640T>G (p.F214V)	5	NA	NA	no	D/D	near active site	likely pathogenic
c.1076C>T (p.A359V)	5	0.03-0.27	1.02×10-8 to 3.95×10-4	no	D/D	alpha-helical bundle	likely pathogenic
c.1019G>T (p.G340V)	5	NA	NA	no	D/D	near active site	likely pathogenic
c.889_891delGAG (p.E297del)	4	NA	NA	no	NA	dimeric interface	likely pathogenic
c.1600G>A (p.E534K)	4	3.6-40.0	0.79 to 2.14×10-12	(Mathur et al 1999)	T/T	surface	likely benign
c.1103A>C (p.Q368P)	4	0.01-0.02	8.06×10-8 to 8.45×10-7	no	D/D	near active site	likely pathogenic

^*The allele count equals the number of unique patients with the variant, all patients were heterozygous for the listed variants. The exception is p.A359V which was found homozygous in one individual.

^¶Expected counts were calculated using the allele frequencies in the ExAC database: (low value) uses overall allele frequency and (high value) uses highest population allele frequency. NA indicates that the allele was not present in the ExAC database.

^¥P-values were calculated using a Fischer's exact test and the same allele frequencies used to generate expected count.

^$Variant prediction; D=deleterious and T=tolerated

^§See Fig. 2