Susceptibility of the interchain peptide of a bromelain inhibitor precursor to the target proteases bromelain, chymotrypsin, and trypsin
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Yoriko Sawano
Abstract
Bromein, a cysteine proteinase inhibitor from pineapple stem, is a unique double-chain inhibitor. The 27.5-kDa precursor protein is processed by the removal of three interchain, two interdomain, and two terminal-flanking peptides, thus resulting in the release of mature isoinhibitors of approximately 6 kDa. To characterize the processing of the interchain peptide Thr15-Ser-Ser-Ser-Asp, we expressed a single-chain precursor with this peptide and monitored proteolytic cleavage by the target proteinase bromelain. By peptide sequencing and mass spectrometric analysis, the initial cleavage was found to occur in vitro between the light-chain and interchain peptides; subsequent trimming formed the terminal-ragged peptides Thr15–Lys60, Ser17–Lys60, Ser18–Lys60, and Asp19–Lys60. However, bromelain did not show any cleavage activity between the interchain and heavy-chain peptides. We also discovered that cleavage between the light-chain and interchain peptides is essential for the single-chain inhibitor to exhibit full inhibitory activity. Notably, the incompletely processed intermediates showed higher inhibitory activity than either the native bromein or the single-chain precursor. Bromein is also known to weakly inhibit the serine proteinases chymotrypsin and trypsin; however, a recombinant single-chain inhibitor with the interchain peptide was no longer able to inhibit these serine proteinases.
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Artikel in diesem Heft
- Saccharomyces cerevisiae translational activator Cbs1p is associated with translationally active mitochondrial ribosomes
- Evolution of vitamin B2 biosynthesis: riboflavin synthase of Arabidopsis thaliana and its inhibition by riboflavin
- Molecular basis of the complex formation between the two calcium-binding proteins S100A8 (MRP8) and S100A9 (MRP14)
- An extracellular carboxylesterase from the basidiomycete Pleurotus sapidus hydrolyses xanthophyll esters
- The composition, structural properties and binding of very-low-density and low-density lipoproteins to the LDL receptor in normo- and hypertriglyceridemia: relation to the apolipoprotein E phenotype
- Adrenodoxin (Adx) and CYP11A1 (P450scc) induce apoptosis by the generation of reactive oxygen species in mitochondria
- Ultraspiracle promotes the nuclear localization of ecdysteroid receptor in mammalian cells
- Polyadenylate polymerase modulations in human epithelioid cervix and breast cancer cell lines, treated with etoposide or cordycepin, follow cell cycle rather than apoptosis induction
- The anti-inflammatory compound curcumin inhibits Neisseria gonorrhoeae-induced NF-κB signaling, release of pro-inflammatory cytokines/chemokines and attenuates adhesion in late infection
- Susceptibility of the interchain peptide of a bromelain inhibitor precursor to the target proteases bromelain, chymotrypsin, and trypsin
- Blocking effect of a biotinylated protease inhibitor on the egress of Plasmodium falciparum merozoites from infected red blood cells
