Fibrolase Trials and Tribulations.docVIP

Toxins 2010, 2, 793-808; doi:10.3390/toxins2040793 OPEN ACCESS toxins ISSN 2072-6651 /journal/toxins Review Fibrolase: Trials and Tribulations Francis S. Markland 1,2,* and Steve Swenson 1,2 1 Department of Biochemistry and Molecular Biology, Cancer Research Laboratory, Keck School of Medicine, University of Southern California, 1303 N. Mission Rd., Los Angeles, CA 90033, USA USC/Norris Comprehensive Cancer Center, Keck School of Medicine, University of Southern California, Los Angeles, CA 90033, USA; E-Mail: sswenson@ 2 * Author to whom correspondence should be addressed; E-Mail: markland@; Tel.: +1-(323) 224-7981; Fax: +1-(323) 224-7679. Received: 11 March 2010; in revised form: 31 March 2010 / Accepted: 19 April 2010 / Published: 20 April 2010 Abstract: Fibrolase is the fibrinolytic enzyme isolated from Agkistrodon contortrix contortrix (southern copperhead snake) venom. The enzyme was purified by a three-step HPLC procedure and was shown to be homogeneous by standard criteria including reverse phase HPLC, molecular sieve chromatography and SDS. The purified enzyme is a zinc metalloproteinase containing one mole of zinc. It is composed of 203 amino acids with a blocked amino-terminus due to cyclization of the terminal Gln residue. Fibrolase shares a significant degree of homology with enzymes of the reprolysin sub-family of metalloproteinases including an active site homology of close to 100%; it is rapidly inhibited by chelating agents such as EDTA, and by alpha2-macroglobulin (???). The enzyme is a direct-acting thrombolytic agent and does not rely on plasminogen for clot dissolution. Fibrolase rapidly cleaves the A(?)-chain of fibrinogen and the B(?)-chain at a slower rate; it has no activity on the ?-chain. The enzyme exhibits the same specificity with fibrin, cleaving the ?-chain more rapidly than the ?-chain. Fibrolase was shown to have very effective thrombolytic