Modulation of the susceptibility of human erythrocytes to snake venom myotoxic phospholipases A2: Role of negatively charged phospholipids as potential membrane binding sites

Cecilia Díaz, Guillermo León, Alexandra Rucavado, Norman Rojas, Alan J. Schroit, José María Gutiérrez

Research output: Contribution to journalArticle

31 Citations (Scopus)

Abstract

Cerrophidion (Bothrops) godmani myotoxins I (CGMT-I) and II (CGMT-II), Asp-49 and Lys-49 phospholipases A2 (PLA2s), which drastically differ in enzymatic activity, were devoid of direct hemolytic effects on erythrocytes (RBC) from different species despite the fact that enzymatically active CGMT-I was able to hydrolyze RBC membrane phospholipids and disrupt liposomes prepared from RBC lipids. Human RBC did not become susceptible to the toxins after treatment with neuraminidase or after altering membrane fluidity with cholesterol or sublytic concentrations of detergent. Unlike normal RBC, significant hemolysis was induced by CGMT-II and another similar Lys-49 isoform, B. asper MT-II (BAMT-II), in RBC enriched with phosphatidylserine (PS). Hemolysis was greater in RBC preincubated with pyridyldithioethylamine (PDA), a potent inhibitor of aminophospholipid transport. RBC enriched with phosphatidic acid (PA) also became susceptible to the myotoxins but was unaffected by PDA. Cells enriched with phosphatidylcholine (PC) remained resistant to the action of the toxins. BAMT-II also induced damage in black lipid membranes prepared with PS but not PC alone. When RBC binding of BAMT-II was measured by enzyme-linked immunosorbent assay, it was observed that PS- and PA-enriched erythrocytes were always able to capture more toxin than normal and PC-enriched RBC. This effect was significantly improved by PDA (in the case of PS) and it was observed either in the presence or in the absence of calcium in the medium. These data suggest that negatively charged lipids in the outer leaflet of cell membranes constitute myotoxic PLA2 binding sites. The scarcity of anionic phospholipids in the outer leaflet of RBC could explain their resistance to the action of these PLA2S.

Original languageEnglish (US)
Pages (from-to)56-64
Number of pages9
JournalArchives of Biochemistry and Biophysics
Volume291
Issue number1
DOIs
StatePublished - 1992

Fingerprint

Snake Venoms
Phospholipases A2
Phosphatidylserines
Membrane Potentials
Phospholipids
Erythrocytes
Phosphatidylcholines
Binding Sites
Modulation
Membranes
Phosphatidic Acids
Hemolysis
Bothrops
Lipids
Immunosorbents
Membrane Fluidity
Fluidity
Neuraminidase
Cell membranes
Membrane Lipids

ASJC Scopus subject areas

  • Biochemistry
  • Biophysics
  • Molecular Biology

Cite this

Modulation of the susceptibility of human erythrocytes to snake venom myotoxic phospholipases A2 : Role of negatively charged phospholipids as potential membrane binding sites. / Díaz, Cecilia; León, Guillermo; Rucavado, Alexandra; Rojas, Norman; Schroit, Alan J.; Gutiérrez, José María.

In: Archives of Biochemistry and Biophysics, Vol. 291, No. 1, 1992, p. 56-64.

Research output: Contribution to journalArticle

Díaz, Cecilia ; León, Guillermo ; Rucavado, Alexandra ; Rojas, Norman ; Schroit, Alan J. ; Gutiérrez, José María. / Modulation of the susceptibility of human erythrocytes to snake venom myotoxic phospholipases A2 : Role of negatively charged phospholipids as potential membrane binding sites. In: Archives of Biochemistry and Biophysics. 1992 ; Vol. 291, No. 1. pp. 56-64.
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