Identification of a precursor processing protease from the spider Cupiennius salei essential for venom neurotoxin maturation

Supporting Information

  • Supplemental Data (.pdf, 7.7 MB) - Supplemental Data Fig. S-1 - S-19, Table S-3 - S-8: Figure S-1. MALDI-TOF spectrum of the purified PQM protease using Sinapic acid as matrix. Figure S-2. MALDI-TOF spectrum of the purified PQM protease using DHAP as matrix. Figure S-3. Fragment ion mass spectrum of the alkylated light chain of the PQM protease. Figure S-4. Mass spectrum of the PQM protease heavy chain. Figure S-5. HCD fragment ion mass spectrum of the disulfide bridge connected peptides GYVNAICLPPGLTNPTGEATVIGWGMLRDDGPISAELR, and KLEPVKGCGK. Figure S-6. EThcD fragment ion mass spectrum of the disulfide bridge connected peptides GYVNAICLPPGLTNPTGEATVIGWGMLRDDGPISAELR, and KLEPVKGCGK Figure S-7. HCD fragment ion mass spectrum of the peptide KLEPVKGCGK generated by EThcD fragmentation of the precursor GYVNAICLPPGLTNPTGEATVIGWGMLRDDGPISAELR, and KLEPVKGCGK (connected through a disulfide bridge). Figure S-8. Comparison of mass spectra of the peptide MQQACGGAILNENWIVTAAHCIDQPINLNNYEIYAGLFSILK with, and without reduced Cys-residues. Figure S-9. HCD fragment ion mass spectrum of the peptide MQQACGGAILNENWIVTAAHCIDQPINLNNYEIYAGLFSILK featuring an intrachain disulfide bridge. Figure S-10. HCD fragment ion mass spectrum of the disulfide bridge connected peptides KIYGDKNSEFDFVQVVPSMLCAGGNGK, and EVQVPIVPWANCK. Figure S-11. HCD fragment ion mass spectrum of the disulfide bridge connected peptides DSCQYDSGGPLFQYDK, and SGGDCGHGPYPGMY. Figure S-12. LC-MS of the synthetic peptide PFFENEQARSC(cam)IPK after overnight incubation with the purified protease. Figure S-13. LC-MS of the synthetic peptide PFLAREQVRKDDKN after overnight incubation with the purified protease. Figure S-14. LC-MS of the synthetic peptide Dabcyl -FYPSQRSETD-Glu(EDANS)-NH2 after overnight incubation with the purified protease. Figure S-15. LC-MS of the synthetic peptide Dabcyl-PFFENEQARKDDKN-Glu(EDANS)-NH2 after overnight incubation with the purified protease. Figure S-16. LC-MS of the synthetic peptide FYPSQRSETDRAKKEL after overnight incubation with the purified protease. Figure S-17. LC-MS of the synthetic peptide WGLEWRNEEAERSPC(cam) after overnight incubation with the purified protease. Figure S-18. LC-MS of the synthetic peptide WGLDWRSEESERSPC(cam) after overnight incubation with the purified protease. Figure S-19. Multiple sequence alignment of homologs of the PQM protease from C. salei (CUPSA). Table S-3. Identified ions from the light chain fragment ion mass spectrum. Table S-4. Identified b- and y- ions from the fragment ion mass spectrum in Figure S-5. Table S-5. Identified b- and y- ions from the fragment ion mass spectrum in Figure S-7. Table S-6. Identified b- and y- ions from the fragment ion mass spectrum in Figure S-9. Table S-7. Identified b- and y- ions from the fragment ion mass spectrum in Figure S-10. Table S-8. Identified b- and y- ions from the fragment ion mass spectrum in Figure S-11.
  • Supplemental Data Table S-2 (.xlsx, 93 KB) - Table S-2. Details of identified proteins / peptides by tandem mass spectrometry of RP-HPLC peak 2
  • Supplemental Data Table S-9 (.xlsx, 325 KB) - Table S-9. Details of identified proteins / peptides by tandem mass spectrometry of the PQM protease containing RP-HPLC peak 3
  • Supplemental Data Table S-1 (.xlsx, 110 KB) - Table S-1. Details of identified proteins / peptides by tandem mass spectrometry of RP-HPLC peak 1