Fig. 5: Sequence conservation of the central tandem repeats in PbINP.

a Comparison of each numbered repeat in PbINP, from the N to C terminus, to the consensus peptide sequence: AGYGSTQTAGEESSLT. Data points in black represent the number of matches to the 16-residue sequence, where a perfect match is 16. The narrow dark blue bar spanning the bottom of the graph represents the central domain. Repeats −3 to −1 compare the last 48 residues of the N-terminal domain to the 16-residue consensus sequence, while repeats 66 and 67 correspond to the first 32 residues of the 41-residue C-terminal domain. Horizontal arrows indicate the repeat region deleted in three different half constructs. b WebLOGO plots⁷³ of the first 54 and last 11 sixteen-residue repeats in PbINP. Each stack’s overall height indicates its degree of sequence conservation, while the height of a letter’s symbol refers to its proportional frequency at that position. c AlphaFold model of the C-terminal region of PbINP. Beta strands are represented as arrows. The central repetitive domain is shown in dark blue, the C-terminal domain in yellow, the R side chains in light blue, and the Y side chains in orange. Repeats 49 to 65 are numbered from the N-terminal end of the central domain. d BINARY nucleation curves for three PbINP constructs in which the front (purple), middle (blue), and back (green) halves of the central repetitive domain were deleted, leaving 33 repeats in each sample. The PstI sites used to delete the back half were present in an earlier version of the synthetic gene and removed repeats 33–64 inclusive. The nucleation curves corresponding to the C-terminal domain deletion (yellow), full-length PbINP (black), and the buffer control (dark blue) are also shown.