Three-dimensional structure of fibrolase, the fibrinolytic enzyme from southern copperhead venom, modeled from the X-ray structure of adamalysin II and atrolysin C.
The fibrinolytic enzyme from southern copperhead snake venom, fibrolase, contains 1 mole of zinc per mole of protein, belongs to the major family of metalloproteinases known as the metzincins, and has been shown to degrade fibrin clots in vitro and in vivo. The purpose of this study was to develop a 3-dimensional model of fibrolase to investigate the geometry of conserved and variable sequences between members of the snake venom metalloproteinases. When compared to atrolysin C (form D) or adamalysin II (metzincins with completely different substrate specificity), fibrolase has approximately 60% overall sequence identity and nearly 100% sequence similarity in the active site. We used the crystal structure of adamalysin II to build a 3-dimensional homology model of fibrolase. Three disulfide bonds were constructed (the highly conserved disulfide bond [118-198] was maintained from the adamalysin II structure and 2 new disulfide bonds were introduced between residues 158-182 and 160-165). We used Sculpt 2.5 and HyperChem 5.0 to "dock" a substrate fragment octapeptide (HTEKLVTS), and a water molecule into the active site cleft. We calculated the differential average homology profile for fibrolase compared to 8 hemorrhagic and 5 nonhemorrhagic metzincins. We then determined the sequence regions that might be responsible for their substrate specificity. Our 3-dimensional homology model shows that the variable sequences lie on the periphery of the identified active site region containing the His triangle; this indicates that substrate specificity may depend on surface residues that are not directly associated with the active site.
Bolger
M B
MB
Department of Pharmaceutical Sciences, University of Southern California, School of Pharmacy, Los Angeles, CA 90089, USA. bolger@usc.edu
Swenson
S
S
Markland
F S
FS
D017836:Agkistrodon; D000818:Animals; D001665:Binding Sites; D018360:Crystallography, X-Ray; D008666:Metalloendopeptidases / Q000737:chemistry*; D008958:Models, Molecular; D009842:Oligopeptides / Q000737:chemistry; D011487:Protein Conformation; D017386:Sequence Homology, Amino Acid; D014757:Viper Venoms / Q000737:chemistry*; D014867:Water / Q000737:chemistry
D009842:Oligopeptides; D014757:Viper Venoms; D014867:Water; D008666:Metalloendopeptidases; C058789:fibrolase; C089232:atrolysin C; C028220:Crotalus adamanteus proteinase II