RT Journal Article
T1 Evaluation of different approaches used to study membrane permeabilization by actinoporins on model lipid vesicles
A1 Palacios Ortega, Juan
A1 Rivera-de-Torre, Esperanza
A1 Gavilanes, José G.
A1 Slotte, J. Peter
A1 Martínez del Pozo, Álvaro
AB Release of aqueous contents from model lipid vesicles has been a standard procedure to evaluate pore formation efficiency by actinoporins, such as sticholysin II (StnII), for the last few decades. However, regardless of the probe of choice, the results reported that StnII action was never able to empty the vesicles completely. This was hard to explain if StnII pores were to be stable and always leaky for the probes used. To address this question, we have used a variety of probes, including rhodamine 6G or Tb3+, to test the permeability of StnII's pores. Our results indicate that calcein was in fact too large to fit through StnII's pores, and that the standard method in the field is actually reporting StnII-induced transient permeation of the membrane rather than the passage of solutes through the stable assembled pores. In order to evaluate the permeability of these structures, we used a dithionite-based assay, which showed that the final pores were in fact open. Thus, our results indicate that the stable actinoporins' pores are open in spite of plateaued classic release curves. Besides the proper pore, the first stages of pore formation would inflict serious damage to living cells as well.
PB Elsevier
SN 0005-2736
YR 2020
FD 2020-04-21
LK https://hdl.handle.net/20.500.14352/6388
UL https://hdl.handle.net/20.500.14352/6388
LA eng
NO Universidad Complutense de Madrid (UCM)/Banco de Santander
NO ISB/ÅA Åbo Akademi University, Turku, Finland
NO Sigrid Jusélius Foundation
NO Jane and Aatos Erkko Foundation
NO Magnus Ehrnrooth Foundation
DS Docta Complutense
RD 7 may 2024