Premium
Stereo‐specific inhibition of sea urchin envelysin (hatching enzyme) by a synthetic autoinhibitor peptide with a cysteine‐switch consensus sequence
Author(s) -
Nomura Kohji,
Suzuki Norio
Publication year - 1993
Publication title -
febs letters
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.593
H-Index - 257
eISSN - 1873-3468
pISSN - 0014-5793
DOI - 10.1016/0014-5793(93)80626-6
Subject(s) - thermolysin , peptide , cysteine , biochemistry , peptide sequence , protease , stereochemistry , cleavage (geology) , consensus sequence , cysteine protease , chemistry , sea urchin , residue (chemistry) , amino acid , enzyme , biology , microbiology and biotechnology , trypsin , paleontology , fracture (geology) , gene
Inhibition of envelysin, a metalloproteinase which dissolves the fertilization envelope of sea urchin embryo, was studied using a synthetic autoinhibitor peptide, Ac‐Pro‐Arg‐Cys‐Gly‐Val‐Pro‐Asp‐Val‐NH 2 , with a ‘cysteine‐switch’ consensus sequence. Although its effect is reversible, the hatching of sea urchin embryos was effectively delayed by 0.5 mM of the peptide. When α 1 ‐proteinase inhibitor was used as the substrate, envelysin was inhibited by the autoinhibitor and an Ala 6 analogue, but not by a d ‐Cys 3 analogue. However, envelysin was weakly inhibited by both d ‐ and l ‐cysteines to the same extent. Snake venom α‐protease exhibited cleavage and inhibition behavior similar to envelysin with a little weaker stereo‐specificity. The results suggest that the coordination of the autoinhibitor Cys residue with the envelysin active site Zn is established only after the amino acid residues on both sides of the Cys residue get into an appropriate interaction with the catalytic site residues, and that the precise orientation of the cysteine SH group is essential. By contrast, thermolysin was weakly inhibited by the three peptide non‐stereo‐specifically. Furthermore, thermolysin cleaved the autoinhibitor at the Cys 3 Gly 4 bond when incubated without substrate.