The purification and partial characterisation of two novel metalloproteinases from the venom of the West African carpet viper, Echis ocellatus

J. M. Howes; Mark Wilkinson; R. D.G. Theakston; G. D. Laing

doi:10.1016/s0041-0101(03)00096-5

The purification and partial characterisation of two novel metalloproteinases from the venom of the West African carpet viper, Echis ocellatus

J. M. Howes, Mark Wilkinson, R. D.G. Theakston, G. D. Laing

Tropical Disease Biology

Research output: Contribution to journal › Article › peer-review

19 Citations (Scopus)

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 metal loproteinase (EoVMP1, p17.0), and a haemorrhagic 56 kDa metalloproteinase (EoVMP2, p15.5). 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. (C) 2003 Elsevier Science Ltd. All rights reserved.

Original language	English
Pages (from-to)	21-27
Number of pages	7
Journal	Toxicon
Volume	42
Issue number	1
DOIs	https://doi.org/10.1016/s0041-0101(03)00096-5
Publication status	Published - 1 Jul 2003

Keywords

Collagenase
Enzyme linked immunosorbant assay
Gelatinase
Haemorrhage
Metalloproteinase
Venom

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10.1016/s0041-0101(03)00096-5

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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 metal loproteinase (EoVMP1, p17.0), and a haemorrhagic 56 kDa metalloproteinase (EoVMP2, p15.5). 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. (C) 2003 Elsevier Science Ltd. All rights reserved.",

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T1 - The purification and partial characterisation of two novel metalloproteinases from the venom of the West African carpet viper, Echis ocellatus

AU - Howes, J. M.

AU - Wilkinson, Mark

AU - Theakston, R. D.G.

AU - Laing, G. D.

PY - 2003/7/1

Y1 - 2003/7/1

N2 - 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 metal loproteinase (EoVMP1, p17.0), and a haemorrhagic 56 kDa metalloproteinase (EoVMP2, p15.5). 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. (C) 2003 Elsevier Science Ltd. All rights reserved.

AB - 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 metal loproteinase (EoVMP1, p17.0), and a haemorrhagic 56 kDa metalloproteinase (EoVMP2, p15.5). 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. (C) 2003 Elsevier Science Ltd. All rights reserved.

KW - Collagenase

KW - Enzyme linked immunosorbant assay

KW - Gelatinase

KW - Haemorrhage

KW - Metalloproteinase

KW - Venom

U2 - 10.1016/s0041-0101(03)00096-5

DO - 10.1016/s0041-0101(03)00096-5

M3 - Article

SN - 0041-0101

VL - 42

SP - 21

EP - 27

JO - Toxicon

JF - Toxicon

IS - 1

ER -

The purification and partial characterisation of two novel metalloproteinases from the venom of the West African carpet viper, Echis ocellatus

Abstract

Keywords

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