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Structural Characterization and Neuromuscular Activity of a New Lys49 Phospholipase A2 Homologous (Bp-12) Isolated from Bothrops pauloensis Snake Venom

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Abstract

Bp-12 was isolated from Bothrops pauloensis snake venom in only one chromatographic step in reverse phase HPLC on μ-Bondapack C-18. The molecular mass of 13,789.56 Da was determined by mass spectrometry. The amino acids composition showed that Bp-12 presented high content of Lys, Tyr, Gly, Pro, and 14 half-Cys residues, typical of a basic PLA2. The sequence of Bp-12 contains 122 amino acid residues: SLFELGKMIL QETGKNPAKS LGAFYCYCGW GSQGQPKDAV DRCCYVHKCC YKKITGCNPK KDRYSYSWKD KTLVCGEDNS CLKELCECDK AVAICLRENL NTYNKKYRYF LKPLCKKADA AC, with a pI value of 8.55 and with a high homology with Lys49 PLA2 from other snake venoms. In mouse phrenic nerve-diaphragm, the time needed for 50% paralysis was: 45 ± 6 min (1.4 μM) and 16 ± 6 min (3.6 μM). Bp-12 can induce indirect and directly blocked evoked twitches, even in the preparations in which Ca2+ is replaced by Sr2+, being the addition of d-tubocurarine required for direct blocking. These results identify Bp-12 as a new member of the Lys49 PLA2 family and shows that this toxin might contribute to the effects of the crude venom on the neuromuscular junction.

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Abbreviations

Asp49:

Aspartate 49

B. pauloensis :

Bothrops pauloensis

Ca2+ :

Calcium ion

HPLC:

High performance liquid chromatography

Lys49:

Lysine 49

MALDI-TOF:

Matrix assisted laser desorption ionisation-time of flight

PLA2 :

Phospholipase A2

Sr2+ :

Strontium ion

TFA:

Trifluoroacetic acid

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Acknowledgments

The authors thank Mr. Paulo A. Baldasso and Gildo Bernardo Leite for technical assistance and Georgina Sucasaca Monzón for some biochemical informations. This work was supported by the National Council for Scientific and Technological Development (CNPq) and is part of Priscila Randazzo-Moura PhD thesis.

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Authors and Affiliations

  1. Pharmacology Department, Medical Science Faculty, State University of Campinas (UNICAMP), Campinas, SP, Brazil

    Priscila Randazzo-Moura & Léa Rodrigues-Simioni

  2. Biochemistry Department, Institute of Biology, State University of Campinas (UNICAMP), Campinas, SP, Brazil

    L. A. Ponce-Soto & Sérgio Marangoni

Authors
  1. Priscila Randazzo-Moura
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  2. L. A. Ponce-Soto
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  3. Léa Rodrigues-Simioni
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  4. Sérgio Marangoni
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Correspondence to Léa Rodrigues-Simioni.

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Randazzo-Moura, P., Ponce-Soto, L.A., Rodrigues-Simioni, L. et al. Structural Characterization and Neuromuscular Activity of a New Lys49 Phospholipase A2 Homologous (Bp-12) Isolated from Bothrops pauloensis Snake Venom. Protein J 27, 355–362 (2008). https://doi.org/10.1007/s10930-008-9144-1

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