Figure 1

Synthetic design and characterization of MBP-CC1. (A) Amino acid composition of CC1. Light blue is soluble protein 2jua and in red is hCLDN1. All chimeric constructs have MBP as N-terminal fusion and a 6xHis tag at the C-terminus. Additionally, we present the full amino acid sequence of the translated CC1. (B) Graphical representation of the chimeric design to produce CC1. (C) Graphical representation of the chimeric design to produce MBP-CC1. The dimensions of relevant axis of MBP-CC1 are presented. MBP, in light blue, also displays, in dark blue, the position and structure of the short linker of four amino acids (Asn, Ala, Ala, Ala). (D) Size-exclusion chromatography of MBP-CC1. The monomeric MBP-CC1 has a molecular weight of 65 kDa. When comparing its elution volume with the protein standards (BioRad Gel Filtration Standard, cat. Number 1511901), MBP-CC1 appears to elute above 670 kDa. Below the x-axis, Vo is void volume, and the numbers correspond to standards of molecular weight. (E) Small-angle X-ray scattering (bioSAXS) of MBP-CC1 performed at 21 °C. Two different orientations of the volumetric data are presented. bioSAXS data collected for MBP-CC1resulted in a radius of gyration (Rg) of 83.1 ± 4.03, and a maximum particle size (Dmax) of 305 Å.