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Purification and Complete Primary Structure of the First PLA2 from Lachesis stenophrys (the Central American Bushmaster) Snake Venom

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Abstract

The first PLA2 (LsPA-1) from L. stenophrys snake venom was purified to homogeneity using three chromatographic steps and had its complete primary structure determined. An average molecular mass of 13,870.3 kDa was determined by mass spectrometry and a 3.3-fold increase in the PLA2 activity was observed for LsPA-1 as compared to the whole venom. Multiple alignment of PLA2 from Lachesis spp. snakes suggested the existence of two geographical clades for this genus in the New World, which is in accordance with morphological, behavioral and mtDNA data obtained by others. Phospholipases A2 from Crotalus spp. snake venoms were similarly distributed into two groups. Intergroup analysis indicated that most amino acid substitutions were observed in the amino- and carboxy-terminal regions of the molecules in each clade. Both regions have been suggested to play important roles in determining the biological properties of PLA2 from snake venoms. The dendogram derived for PLA2 from Lachesis and Crotalus snakes highlighted the phylogenetic relationships between these two genera in the New World.

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Abbreviations

FP3:

Pool 3 from filtration chromatography

LmPLA2I and LmPLA2II:

Isoforms of PLA2 from Lachesis muta snake venom

LmTx1 and LmTx2:

Isoforms of PLA2from Lachesis muta muta snake venom

LsPA-1:

PLA2 from Lachesis stenophrys snake venom

Maldi:

Matrix assisted laser desorption ionization

MtDNA:

Mitochondrial DNA

MQP3:

Pool 3 from MonoQ chromatography

Nm:

Nanometers

PBS:

Phosphate buffered saline

PLA2 :

Phospholipase A2

SDS-PAGE:

Polyacrylamide gel electrophoresis with SDS

TFA:

Trifluoroacetic acid

TOF:

Time-of-flight

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Acknowledgements

We thank Dr. Márcia Helena Borges (FUNED) for helping in the amino acid sequencing and Dr. Adriano Pimenta and Mr. Rodrigo Novaes Ferreira for the mass spectrometry. This work was financially supported by the following Brazilian agencies: FAPEMIG (Fundação de Amparo à Pesquisa do Estado de Minas Gerais) and CNPq (Conselho Nacional de Desenvolvimento Científico e Tecnológico).

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Authors and Affiliations

  1. Laboratory of Molecular and Cell Biology, Division of Biomedical Sciences, Fundação Ezequiel Dias (FUNED), Rua Conde Pereira Carneiro 80, CEP 30510-010, Belo Horizonte, MG, Brazil

    Eduardo Borges de Assis, Maria Inácia Estevão-Costa, Ana do Carmo Valentim, Aristeu Silva-Neto, Maurício Alvarenga Mudado & Michael Richardson

  2. Serpentarium, Division of Immunobiologicals, Fundação Ezequiel Dias (FUNED), Rua Conde Pereira Carneiro 80, CEP 30510-010, Belo Horizonte, MG, Brazil

    Giselle Agostini Cotta

  3. Fundação Ezequiel Dias/DPD, Rua Conde Pereira Carneiro 80, CEP 30510-010, Belo Horizonte, MG, Brazil

    Consuelo Latorre Fortes-Dias

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  1. Eduardo Borges de Assis
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  2. Maria Inácia Estevão-Costa
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  3. Ana do Carmo Valentim
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  4. Aristeu Silva-Neto
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  5. Giselle Agostini Cotta
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  6. Maurício Alvarenga Mudado
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  7. Michael Richardson
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  8. Consuelo Latorre Fortes-Dias
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Correspondence to Consuelo Latorre Fortes-Dias.

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de Assis, E.B., Estevão-Costa, M.I., do Carmo Valentim, A. et al. Purification and Complete Primary Structure of the First PLA2 from Lachesis stenophrys (the Central American Bushmaster) Snake Venom. Protein J 27, 327–333 (2008). https://doi.org/10.1007/s10930-008-9141-4

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  • DOI: https://doi.org/10.1007/s10930-008-9141-4

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