Characterization of a fibrinogen-clotting enzyme from Trimeresurus stejnegeri venom, and comparative study with other venom proteases

Trimeresurus stejnegeri venom, which contains TSV-PA (a specific plasminogen activator sharing 60-70% sequence homology with venom fibrinogen-clotting enzymes), also possesses fibrinogen-clotting activity in vitro. A fibrinogen-clotting enzyme (stejnobin) has been purified to homogeneity by gel filtration and ion-exchange chromatography on a Mono-Q column. It is a single-chain glycoprotein with a mol. wt of 44,000. The NH2-terminal amino acid sequence of stejnobin shows great homology with venom fibrinogen-clotting enzymes and TSV-PA. Like TSV-PA, stejnobin was able to hydrolyse several chromogenic substrates. Comparative study of substrate specificities of stejnobin and other venom proteases purified in our laboratory was carried out on five chromogenic substrates. Stejnobin clotted human fibrinogen with a specific activity of 122 NIH thrombin-equivalent units/mg protein. However, stejnobin did not act on other blood coagulation factors, such as factor X, prothrombin and plasminogen. Diisopropyl fluorophosphate and phenylmethanesulfonyl fluoride inhibited its activity, whereas ethylenediamine tetracetic acid had no effect on it, indicating that it is a serine protease. Although stejnobin showed strong immunological cross-reaction with polyclonal antibodies raised against TSV-PA, it was interesting to observe that, unlike the case of TSV-PA, these antibodies did not inhibit the amidolytic and fibrinogen-clotting activities of stejnobin. (C) 1998 Elsevier Science Ltd. All rights reserved.

Production cross-section of Ra-230 in the reaction of 60 MeV/u O-18 ions with U-238

A thick natural uranium target was bombarded with a 60 MeV/u O-18 beam. The neutron-rich isotope Ra-230 as the target residue was produced through the multinucleon transfer reaction (U-238-4p-4n). The barium and radium fraction as BaCl2 precipitate were radiochemically separated first from the mixture of uranium and reaction products. Then, the radium fraction was separated from BaCl2 precipitate by using cation exchange technique. The gamma-ray spectra of the Ra fraction were measured using an HPGe detector. The production cross sections of Ra-230 were obtained by a combination of the radiochemical separation technique and off-line gamma-ray spectroscopy. The cross section of Ra-230 has been determined to be 66 +/- 20 mu b.