Interaction of viscotoxins A3 and B with membrane model systems: implications to their mechanism of action.
Viscotoxins are small proteins that are thought to interact with biomembranes, displaying different toxic activities against a varied number of cell types, being viscotoxin A(3) (VtA(3)) the most cytotoxic whereas viscotoxin B (VtB) is the less potent. By using infrared and fluorescence spectroscopies, we have studied the interaction of VtA(3) and VtB, both wild and reduced ones, with model membranes containing negatively charged phospholipids. Both VtA(3) and VtB present a high conformational stability, and a similar conformation both in solution and when bound to membranes. In solution, the infrared spectra of the reduced proteins show an increase in bandwidth compared to the nonreduced ones indicating a greater flexibility. VtA(3) and VtB bind with high affinity to membranes containing negatively charged phospholipids and are motional restricted, their binding being dependent on phospholipid composition. Whereas nonreduced proteins maintain their structure when bound to membranes, reduced ones aggregate. Furthermore, leakage experiments show that wild proteins were capable of disrupting membranes whereas reduced proteins were not. The effect of VtA(3) and VtB on membranes having different phospholipid composition is diverse, affecting the cooperativity and fluidity of the membranes. Viscotoxins interact with membranes in a complex way, most likely organizing themselves at the surface inducing the appearance of defects that lead to the destabilization and disruption of the membrane bilayer.
Giudici
Marcela
M
Instituto de Biología Molecular y Celular, Universidad Miguel Hernández, Campus de Elche, E-03202 Elche-Alicante, Spain.
Pascual
Roberto
R
de la Canal
Laura
L
Pfüller
Karola
K
Pfüller
Uwe
U
Villalaín
José
J
D016880:Anisotropy; D004347:Drug Interactions; D008051:Lipid Bilayers / Q000737:chemistry*; D046911:Macromolecular Substances; D008560:Membrane Fluidity; D008567:Membranes, Artificial; D010539:Permeability; D010743:Phospholipids / Q000737:chemistry*; D010940:Plant Proteins / Q000737:chemistry*; D016062:Porosity; D011485:Protein Binding; D011487:Protein Conformation; D013050:Spectrometry, Fluorescence; D013055:Spectrophotometry, Infrared; D013696:Temperature; D013844:Thiazoles; D053778:Thiazolidines
D008051:Lipid Bilayers; D046911:Macromolecular Substances; D008567:Membranes, Artificial; D010743:Phospholipids; D010940:Plant Proteins; D013844:Thiazoles; D053778:Thiazolidines; C485282:viscotoxin B, Viscum album; C485281:viscotoxin A3, Viscum album; C018578:letosteine