The mature bovine cathepsin C (CC) molecule is composed of four identical monomers, each proteolytically processed into three chains. Five intrachain disulfides and three nonpaired cysteine residues per monomer were identified. Beside catalytic Cys234 in the active site, free-thiol Cys331 and Cys424 were characterized. Cys424 can be classified as inaccessible buried residue. Selective modification of Cys331 results in dissociation of native CC tetramer into dimers. The 3D homology-based model of the CC catalytic core suggests that Cys331 becomes exposed as the activation peptide is removed during procathepsin C activation. The model further shows that exposed Cys331 is surrounded by a surface hydrophobic cluster, unique to CC, forming a dimer-dimer interaction interface. Substrate/inhibitor recognition of the active site in the CC dimer differs significantly from that in the native tetramer. Taken together, a mechanism is proposed that assumes that the CC tetramer formation results in a site-specific occlusion of endopeptidase-like active site cleft of each CC monomeric unit. Thus, tetramerization provides for the structural basis of the dipeptidyl peptidase activity of CC through a substrate access-limiting mechanism different from those found in homologous monomeric exopeptidases cathepsin H and B. In conclusion, the mechanism of tetramer formation as well as specific posttranslational processing segregates CC in the family of papain proteases.
- MeSH
- aktivace enzymů MeSH
- cystein chemie MeSH
- disulfidy chemie MeSH
- kathepsin C chemie izolace a purifikace metabolismus MeSH
- konformace proteinů MeSH
- lysin chemie MeSH
- molekulární sekvence - údaje MeSH
- molekulová hmotnost MeSH
- peptidové fragmenty MeSH
- posttranslační úpravy proteinů MeSH
- sbalování proteinů MeSH
- sekvence aminokyselin MeSH
- skot MeSH
- slezina enzymologie MeSH
- vysokoúčinná kapalinová chromatografie MeSH
- zvířata MeSH
- Check Tag
- skot MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- Názvy látek
- cystein MeSH
- disulfidy MeSH
- kathepsin C MeSH
- lysin MeSH
- peptidové fragmenty MeSH
