Figure 1

Domain arrangement of proDerp1αS construct, along with SDS-PAGE, Western blot and mass spectrometry analysis of the purified immunotoxin. (A) Schematic representation of proDerp1αS cDNA, highlighting its structural and functional motifs, alongside its theoretical molecular mass. 3D-model structure of proDerp1αS is included as Figure S1. (B) Coomassie blue stained SDS-PAGE (CBS) and Western blot analysis using rabbit anti-α-sarcin (left) and anti-α-Der p 1 antisera (right). CBS protein molecular weight standards correspond to Bio-Rad Unstained SDS-PAGE low range Standards; while the prestained Bio-Rad Precision. Plus Dual Color Standards (Bio-Rad, Hercules, CA, USA) were used for Western blot. Images correspond to full-length gels and blots acquired and analyzed using the Gel Doc XR Imaging System and Quantity One 1-D analysis sofware (BioRad) or ChemiDoc-It (UVP) and VisionWorks LS, respectively.Full-length blots/gels are presented in Supplementary Figure S2. (C) Mass spectrometry analysis spectrum showing the corresponding peak to proDerp1αS construct. The difference between theoretical and empirical masses agrees with the four N-terminal extra amino acids of α-factor secretion signal that could remain after its processing by kex2 protease.