Characterisation of a purified phospholipase A2 from the venom of the Papuan black snake (Pseudechis papuanus)

Gavin D. Laing; Aura S. Kamiguti; Mark Wilkinson; Gordon M. Lowe; R. David G. Theakston

doi:10.1016/0167-4838(95)00081-5

Characterisation of a purified phospholipase A2 from the venom of the Papuan black snake (Pseudechis papuanus)

Gavin D. Laing, Aura S. Kamiguti, Mark Wilkinson, Gordon M. Lowe, R. David G. Theakston

Liverpool School of Tropical Medicine

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

23 Citations (Scopus)

Abstract

A neutral phospholipase A2 (PLA2) was separated from Pseudechis papuanus venom by a two-stage FPLC procedure of cation exchange and phenyl-Superose chromatography. It had a molecular mass of 15 kDa and a lower LD50 value than a co-separated haemorrhagic fraction, indicating a higher lethal potency. In vitro tests confirmed the powerful inhibition of platelet aggregation by the PLA2 and strong anticoagulant activity initially observed with whole venom. Ultrastructural studies showed that platelets lost their discoid shape and developed membranous projections with a general decrease in electron-density of the cytosol and disruption of the microfilaments following incubation with the enzyme. Amino acid sequence analysis of the N-terminus and some internal peptides demonstrated a high degree of homology with PLA2s from other Pseudechis venoms. Our results indicate that this fraction is the main agent responsible for the haemostatic disorders in envenomed patients.

Original language	English
Pages (from-to)	137-143
Number of pages	7
Journal	Biochimica Et Biophysica Acta-Proteins and Proteomics
Volume	1250
Issue number	2
DOIs	https://doi.org/10.1016/0167-4838(95)00081-5
Publication status	Published - 19 Jul 1995

Keywords

Anticoagulant
Platelet inhibitor
Venom PLA

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10.1016/0167-4838(95)00081-5

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title = "Characterisation of a purified phospholipase A2 from the venom of the Papuan black snake (Pseudechis papuanus)",

abstract = "A neutral phospholipase A2 (PLA2) was separated from Pseudechis papuanus venom by a two-stage FPLC procedure of cation exchange and phenyl-Superose chromatography. It had a molecular mass of 15 kDa and a lower LD50 value than a co-separated haemorrhagic fraction, indicating a higher lethal potency. In vitro tests confirmed the powerful inhibition of platelet aggregation by the PLA2 and strong anticoagulant activity initially observed with whole venom. Ultrastructural studies showed that platelets lost their discoid shape and developed membranous projections with a general decrease in electron-density of the cytosol and disruption of the microfilaments following incubation with the enzyme. Amino acid sequence analysis of the N-terminus and some internal peptides demonstrated a high degree of homology with PLA2s from other Pseudechis venoms. Our results indicate that this fraction is the main agent responsible for the haemostatic disorders in envenomed patients.",

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doi = "10.1016/0167-4838(95)00081-5",

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T1 - Characterisation of a purified phospholipase A2 from the venom of the Papuan black snake (Pseudechis papuanus)

AU - Laing, Gavin D.

AU - Kamiguti, Aura S.

AU - Wilkinson, Mark

AU - Lowe, Gordon M.

AU - Theakston, R. David G.

PY - 1995/7/19

Y1 - 1995/7/19

N2 - A neutral phospholipase A2 (PLA2) was separated from Pseudechis papuanus venom by a two-stage FPLC procedure of cation exchange and phenyl-Superose chromatography. It had a molecular mass of 15 kDa and a lower LD50 value than a co-separated haemorrhagic fraction, indicating a higher lethal potency. In vitro tests confirmed the powerful inhibition of platelet aggregation by the PLA2 and strong anticoagulant activity initially observed with whole venom. Ultrastructural studies showed that platelets lost their discoid shape and developed membranous projections with a general decrease in electron-density of the cytosol and disruption of the microfilaments following incubation with the enzyme. Amino acid sequence analysis of the N-terminus and some internal peptides demonstrated a high degree of homology with PLA2s from other Pseudechis venoms. Our results indicate that this fraction is the main agent responsible for the haemostatic disorders in envenomed patients.

AB - A neutral phospholipase A2 (PLA2) was separated from Pseudechis papuanus venom by a two-stage FPLC procedure of cation exchange and phenyl-Superose chromatography. It had a molecular mass of 15 kDa and a lower LD50 value than a co-separated haemorrhagic fraction, indicating a higher lethal potency. In vitro tests confirmed the powerful inhibition of platelet aggregation by the PLA2 and strong anticoagulant activity initially observed with whole venom. Ultrastructural studies showed that platelets lost their discoid shape and developed membranous projections with a general decrease in electron-density of the cytosol and disruption of the microfilaments following incubation with the enzyme. Amino acid sequence analysis of the N-terminus and some internal peptides demonstrated a high degree of homology with PLA2s from other Pseudechis venoms. Our results indicate that this fraction is the main agent responsible for the haemostatic disorders in envenomed patients.

KW - Anticoagulant

KW - Platelet inhibitor

KW - Venom PLA

U2 - 10.1016/0167-4838(95)00081-5

DO - 10.1016/0167-4838(95)00081-5

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JO - Biochimica Et Biophysica Acta-Proteins and Proteomics

JF - Biochimica Et Biophysica Acta-Proteins and Proteomics

IS - 2

ER -

Characterisation of a purified phospholipase A2 from the venom of the Papuan black snake (Pseudechis papuanus)

Abstract

Keywords

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