title: The purification and partial characterisation of two novel metalloproteinases from the venom of the West African carpet viper, Echis ocellatus.
abstract: Separation of previously uncharacterised Echis ocellatus venom by phenyl-Superose FPLC (Fast Liquid Protein Chromatography) yielded eight protein fractions. Three of these displayed high proteolytic activity when assayed by in vivo and in vitro assays (including enzyme linked immunosorbant assay), and were further separated using Superdex 75 and Mono-Q FPLC. This resulted in the purification of a non-haemorrhagic 24 kDa metalloproteinase (EoVMP1, pI 7.0), and a haemorrhagic 56 kDa metalloproteinase (EoVMP2, pI 5.5). Following tryptic digest, short amino acid sequences of EoVMP1 and EoVMP2 were obtained using Edman degradation. Both sequences displayed homology when aligned with existing snake venom metalloproteinases (SVMPs). The strong homology observed among previously well-characterised SVMPs suggests that principles governing the interaction of substrates and inhibitors are likely to be similar for EoVMP1, EoVMP2 and all members of the reprolysin family.
authors.affiliation: Venom Research Unit, Liverpool School of Tropical Medicine, Pembroke Place, Liverpool L3 5QA, UK.
authors.forename: J-M
authors.initials: JM
authors.lastname: Howes
authors.forename: M C
authors.initials: MC
authors.lastname: Wilkinson
authors.forename: R D G
authors.initials: RD
authors.lastname: Theakston
authors.forename: G D
authors.initials: GD
authors.lastname: Laing
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D002853:Chromatography, Liquid; D024141:Collagen Type IV / Q000187:drug effects / Q000378:metabolism; D004797:Enzyme-Linked Immunosorbent Assay; D006470:Hemorrhage / Q000139:chemically induced / Q000473:pathology; D008297:Male; D008666:Metalloendopeptidases / Q000302:isolation & purification* / Q000494:pharmacology; D051379:Mice; D008815:Mice, Inbred Strains; D008969:Molecular Sequence Data; D016415:Sequence Alignment; D014757:Viper Venoms / Q000737:chemistry*; D017819:Viperidae
chemical_list: D024141:Collagen Type IV; D014757:Viper Venoms; D008666:Metalloendopeptidases