Table 1.
Rs4751440 (L6-NPS) exhibits a high frequency in Europeans based on Exome Sequencing Project.
Table 2.
Rs4751440 (L6-NPS) exhibits a high frequency in Europeans based on Exome Sequencing Project based on Hapmap project.
Figure 1.
Origination of the L6-NPS variant.
(A) Haplotype distribution of the genomic region surrounding SNP rs4751440 based on 20 SNPs covering about 14 kb. The eight most common haplotypes are shown in the left inset, and the remaining low-frequency haplotypes are grouped as “Other.” The chimpanzee allele was assumed to be the ancestral one. The rs4751440 is shown in red whereas all other alleles are in dark blue. (B) The approximate posterior probability density for the geographic origin of L6-NPS obtained by the ABC simulation. The heat map was generated using 2D kernel density estimation of the latitude and longitude coordinates from the top ranked 5,000 of 3,000,000 simulations. Red color represents the highest probability, and blue the lowest.
Figure 2.
Sequence comparison and lower NPSR receptor signaling ability of the L6-NPS variant.
(A) Alignment of NPS among diverse vertebrate species. Sequences were aligned by the ClustalW program [75]. Stars indicate identical amino acids and double dots (:) indicate conserved amino acids. The predicted convertase cleavage sites (KR) were underlined and the 6th amino acid “V” or “L” in the mature peptide region are shown in bold. (B) Comparison of NPS and L6-NPS signaling based on the CRE-luciferase assay. (C) Comparison of NPS and L6-NPS signaling based on the NFAT luciferase assay. HEK293T cells seeded in 24-well plates were co-transfected with different luciferase reporters (50 ng), the pSV-β-Gal plasmid (5 ng), and the NPSR receptor plasmid (50 ng). After 36 h, cells were cultured in serum-free media for another 18 h with increasing doses of the various NPS peptides. EC50 values were analyzed using Graphpad Prism 5.0.
Figure 3.
Signaling abilities of different NPS mutants.
(A) Stimulation of CRE luciferase activity mediated by NPSR in response to increasing dosages of various NPS mutants. (B) Stimulation of NFAT luciferase activity mediated by NPSR in response to increasing dosages of various NPS mutants. EC50 vales were analyzed using Graphpad Prism 5.0.