Antiplasmin cleaving enzyme

An inhibitor of antiplasmin cleaving enzyme which is effective in binding to or blocking the α2-antiplasmin binding site, or α2-antiplasmin pro-asn or cleaving site of antiplasmin cleaving enzyme.

A method for identifying an enzyme inhibitor, comprising:

• combining a dimeric molecule having α2-antiplasmin cleaving enzymatic activity, said dimeric molecule having a molecular weight of about 180 kilodaltons as determined by SDS-PAGE, a substrate for said molecule, and a test substance to be tested as an enzyme inhibitor;
• and
• determining activity of the dimeric molecule on the substrate, wherein a decrease in activity of the dimeric molecule when the test substance is present indicates that said test substance is an inhibitor.

A monoclonal antibody raised against α2-antiplasmin cleaving enzyme which binds to an α2-antiplasmin binding portion of the α2-antiplasmin cleaving enzyme.

A method of screening a subject for risk for atherosclerosis or complications of atherosclerosis or for diseases related to fibrin deposition, comprising:

• determining the identity of the amino acid at position 6 in α2-antiplasmin of the subject.

A method of inhibiting digestion by plasmin in a subject in need of such therapy, comprising:

• administering to the subject a therapeutically effective quantity of α2-antiplasmin cleaving enzyme effective in enhancing the availability of soluble activated α2-antiplasmin in the subject's plasma, the α2-antiplasmin cleaving enzyme comprising a protein having a molecular weight of about 180 kDa in a dimeric form as determined by SDS PAGE, wherein each subunit of the dimeric form has a molecular weight of about 97 kDa as determined by SDS PAGE, the protein comprising an N-terminal sequence comprising SEQ ID NO:1 and internal sequences comprising SEQ ID NO:4, SEQ ID NO:5, SEQ ID NO:6, SEQ ID NO:7, and SEQ ID NO:8, and wherein the enzyme cleaves precursor α2-antiplasmin at the pro12-asn13 bond.

A method of producing activated α2-antiplasmin, in vitro, comprising:

• combining a quantity of precursor α2-antiplasmin with a quantity of α2-antiplasmin cleaving enzyme under conditions suitable for cleavage of the precursor α2-antiplasmin by the α2-antiplasmin cleaving enzyme, the α2-antiplasmin cleaving enzyme comprising:
• a protein having a molecular weight of about 180 kDa in a dimeric form as determined by SDS PAGE, wherein each subunit of the dimeric form has a molecular weight of about 97 kDa as determined by SDS PAGE, the protein comprising an N-terminal sequence comprising SEQ ID NO:1 and internal sequences comprising SEQ ID NO:4, SEQ ID NO:5, SEQ ID NO:6, SEQ ID NO:7, and SEQ ID NO:8, and wherein the enzyme cleaves precursor α2-antiplasmin at the pro12-asn13 bond.

A method of enhancing fibrin digestion in vivo, comprising providing to a subject in need of clot digestion or clot prevention, simultaneously or in sequence, a quantity of plasminogen activator and an inhibitor of antiplasmin cleaving enzyme, wherein the quantity of plasminogen activator is less than the amount provided in standard therapeutic protocol absent the inhibitor of antiplasmin cleaving enzyme.