Purification and characterization of a plasmin-like protease from Tenodera
sinensis (Chinese mantis).
Hahn BS, Cho SY, Ahn MY, Kim YS.
Abstract
A novel type of protease (mantis egg fibrinolytic enzyme, MEF-2) was
isolated from the egg cases of Tenodera sinensis. The protease was homogeneous
by SDS-PAGE and its apparent molecular mass was 32,900 Da. The amino acids in
the N-terminal region were Ile-Val-Gly-Gly-Glu-Glu-Ala-Val-Ala-Gly-Asp-Phe-Pro-Ile-Val-Ser-Leu-Gln-Glu.
The enzyme was inhibited by PMSF, TLCK, aprotinin, benzamidine, soybean trypsin
inhibitor and also slightly by elastatinal, EDTA, EGTA, cysteine and beta-mercaptoethanol,
but TPCK, iodoacetate and E-64 did not affect the activity. MEF-2 was not sensitive
to alpha(1)-antitrypsin but antithrombin III and alpha(2)-antiplasmin inhibited
the enzyme. MEF-2 preferentially cleaved the oxidized B-chain of insulin between
Arg(22) and Gly(23). Among chromogenic protease substrates, the most susceptible
to MEF-2 hydrolysis was benzoyl-Phe-Val-Arg-p-nitroanilide with maximal activity
at 30 degrees C and pH 5.0. These results indicate that MEF-2 belongs to the
trypsin family. Upon incubation of crosslinked fibrin with MEF-2, a steady increase
of D-dimer suggests that the enzyme has a strong fibrinolytic activity. In conclusion,
MEF-2 is a new type of proteolytic enzyme and has some potential for practical
application in fibrinolysis.
PMID: 11267896 Insect Biochem Mol Biol. 2001 Apr 27;31(6-7):573-81. ncbi.nlm.nih.gov