title: Cleavage specificity of porcine follipsin.
abstract: Follipsin purified from the follicular fluid of porcine ovaries was studied for its specificity against various synthetic and peptide substrates. The enzyme cleaved only by an endopeptidase activity at the amide and peptide bonds of Arg-X, indicating strict specificity of the S1 pocket for arginine. The specificity for pocket S2 appears to favor either hydrophobic or basic side chains. A 10-residue peptide containing a portion of the activation site of human tissue plasminogen activator was synthesized and tested with the enzyme. The peptide was cleaved by follipsin at the Arg-Ile bond, as expected from the specificity deduced above. Furthermore, the enzyme successfully converted single-chain precursor tissue plasminogen activator (sctPA) to its active, two-chain form by cleaving the corresponding peptide bond. Comparison of the rates of single-chain precursor tissue plasminogen activator activation and tissue plasminogen activator peptide hydrolysis revealed that the former is a more efficient substrate than the latter.
authors.affiliation: Division of Biological Sciences, Graduate School of Science, Hokkaido University, Sapporo, Japan.
authors.forename: J
authors.initials: J
authors.lastname: Ohnishi
authors.forename: T
authors.initials: T
authors.lastname: Kihara
authors.forename: T
authors.initials: T
authors.lastname: Hamabata
authors.forename: K
authors.initials: K
authors.lastname: Takahashi
authors.forename: T
authors.initials: T
authors.lastname: Takahashi
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D004789:Enzyme Activation; D005260:Female; D006868:Hydrolysis; D008969:Molecular Sequence Data; D010053:Ovary / Q000201:enzymology; D011994:Recombinant Proteins / Q000378:metabolism; D012697:Serine Endopeptidases / Q000378:metabolism*; D013379:Substrate Specificity; D013552:Swine; D014568:Urokinase-Type Plasminogen Activator / Q000378:metabolism
chemical_list: D011994:Recombinant Proteins; D012697:Serine Endopeptidases; C088241:follipsin; D014568:Urokinase-Type Plasminogen Activator; C067551:saruplase