Figure 4.

Intrepicalcin sequence alignment and bioinformatics (A) Sequence alignment of the Intrepicalcin (Genbank accession: JZ818387), with the sequences of other eight calcins: Vejocalcin (Xiao et al., 2016), Maurocalcin (UniProt: P60254), Hemicalcin (Shahbazzadeh et al., 2007), Hadrucalcin (UniProt: B8QG00), Opicalcin₁ (UniProt: P60252), Opicalcin₂ (UniProt: P60253), Urocalcin (UniProt: AGA82762) and Imperacalcin (UniProt: P59868). The identity values to Intrepicalcin are shown in the right side. Positively-charged residues Lysine (K) and Arginine (R), negatively charged residues Aspartic acid (D) and Glutamic acid (E), and the disulfide bond-forming Cysteine (C) are highlighted by the colors blue, red and grey, respectively. Columns with identical residue are marked by asterisk while columns with only one difference are marked by colon in the bottom. Three pairs of highly conserved disulfide bridges (Cys³-Cys¹⁷, Cys¹⁰-Cys²¹ and Cys¹⁶-Cys³²) and the residues forming part of β-strands appear connected to form an ICK motif. (B) Net charge vs. pH plot and Hydrophobicity plot of Intrepicalcin. (C) Three dimensional modeling of Intrepicalcin simulated by Swiss-PdbViewer 4.1.0 and viewed by Discovery Studio 4.0 according to the three-dimensional structure of Imperacalcin resolved by ¹H-NMR (Lee et al., 2004). The solid ribbon with disulfide bridge is located in the left and the charged CPK model with “frontal” side (middle) and “dorsal” side (right) are also displayed. Positively charged residues (Lys and Arg), negatively charged residues (Asp and Glu), and neutral residues are colored by blue, red and grey, respectively.