PMID: 3214172Dec 1, 1988Paper

Multicatalytic proteinase in fish muscle

Archives of Biochemistry and Biophysics
E J FolcoJ J Sanchez

Abstract

Proteinase II, a high-molecular-mass proteinase previously identified in white croaker skeletal muscle, was purified to apparent homogeneity by DEAE-Sephacel, phenyl-Sepharose CL 4B, and Sephacryl S-300 chromatographies. Under denaturing conditions, the enzyme dissociated into a cluster of subunits with Mr ranging from 18,000 to 26,000 and a large subunit with a Mr 60,000. The proteinase was able to hydrolyze N-terminal-blocked 4-methyl-7-coumarylamide substrates having either an aromatic amino acid (chymotrypsin-like activity) or an arginine residue (trypsin-like activity) adjacent to the fluorogenic group. The trypsin-like activity of the enzyme was inhibited by fatty acids and sodium dodecyl sulfate, whereas the chymotrypsin-like activity was stimulated by those compounds but inhibited by nonionic and cationic detergents. Several thiol reagents inhibited both proteinase II activities. However, leupeptin and Cu2+ strongly inhibited its trypsin-like activity but only slightly affected its chymotrypsin-like activity. Dithiothreitol stimulated both activities, but at different extents and in different concentration ranges. These results suggest that the enzyme is multicatalytic, having at least two different active sites.

References

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Citations

Jan 1, 1995·Molecular Biology Reports·B Dahlmann, L Kuehn
Jan 1, 1995·Molecular Biology Reports·J J SánchezC A Casalongué
Jun 1, 1992·Comparative Biochemistry and Physiology. B, Comparative Biochemistry·L BusconiJ J Sanchez
Aug 1, 1995·Comparative Biochemistry and Physiology. Part B, Biochemistry & Molecular Biology·I Stoknes, T Rustad
Dec 31, 2003·Comparative Biochemistry and Physiology. Part B, Biochemistry & Molecular Biology·Makoto OhkuboTadashi Ishihara
Feb 8, 2005·Comparative Biochemistry and Physiology. Part B, Biochemistry & Molecular Biology·Adele R ThomasRyno J Naudé
Dec 8, 2004·Comparative Biochemistry and Physiology. Part B, Biochemistry & Molecular Biology·Makoto OhkuboTadashi Ishihara
Jun 9, 2005·Comparative Biochemistry and Physiology. Part B, Biochemistry & Molecular Biology·Makoto OhkuboFutoshi Aranishi
Aug 15, 1991·Archives of Biochemistry and Biophysics·C B MartoneJ J Sánchez
Jul 14, 1989·Biochemical and Biophysical Research Communications·Y SaitohS Ishii
Feb 14, 1991·Biochemical and Biophysical Research Communications·I OhkuboM Sasaki

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