Infrared spectroscopy study on the conformational changes leading to pore formation of the toxin sticholysin II.
The structure of the actinoporin sticholysin II (StnII) in the pore state was investigated by Fourier transform infrared spectroscopy in the attenuated total reflection configuration. 1-Palmitoyl-2-oleoyl-sn-glycero-3-phosphocholine/cholesterol unilamellar vesicles were employed. The alpha-helix content increases in approximately 30% upon lipid binding, which agrees with an extension of eight or nine residues at the N-terminal helix. Furthermore, analyses of dichroic spectra show that the extended N-terminal helix would have a 31 degrees tilt with respect to the membrane normal. The orientation of the central beta-sandwich was also estimated. In addition, it was detected that StnII alters the orientation of the lipid acyl chains. (1)H/(2)H exchange experiments sustain a mainly superficial interaction between StnII and the membrane, with no protection of the beta-sandwich. The implications of the results in the mechanism of pore formation are discussed.
Alegre-Cebollada
Jorge
J
Departamento de Bioquímica y Biología Molecular I, Facultad de Ciencias Químicas, Universidad Complutense, 28040 Madrid, Spain. alegre@bbm1.ucm.es
Martínez del Pozo
Alvaro
A
Gavilanes
José G
JG
Goormaghtigh
Erik
E
D000818:Animals; D002784:Cholesterol / Q000378:metabolism; D003064:Cnidarian Venoms / Q000737:chemistry* / Q000378:metabolism; D008567:Membranes, Artificial; D010713:Phosphatidylcholines / Q000378:metabolism; D052899:Pore Forming Cytotoxic Proteins / Q000737:chemistry* / Q000378:metabolism; D011487:Protein Conformation; D017433:Protein Structure, Secondary; D012615:Sea Anemones; D013055:Spectrophotometry, Infrared; D014118:Toxins, Biological / Q000737:chemistry* / Q000378:metabolism
D003064:Cnidarian Venoms; D008567:Membranes, Artificial; D010713:Phosphatidylcholines; D052899:Pore Forming Cytotoxic Proteins; D014118:Toxins, Biological; C404794:sticholysin II; D002784:Cholesterol; C028694:1-palmitoyl-2-oleoylphosphatidylcholine