Fluorescence spectroscopic study of the interaction of saporin with phospholipid vesicles.
Saporin-S6 (SO-6) is a type-1 ribosome-inactivating protein purified from the seeds of Saponaria officinalis. The fluorescence characteristics of SO-6 were studied in the presence and absence of phospholipids. The interaction of SO-6 with DMPG or DMPC vesicles results in a decrease in the fluorescence emission intensity of tryptophan without any shift in the emission maximal wavelength. The results of fluorescence titration indicate that DMPG/SO-6 saturation molar ratio is 100: 1, but the binding of DMPC with SO-6 does not reach a saturating plot. A shielding of the tryptophan fluorescence from quenching by acrylamide on interaction with the phospholipids was observed, and this shielding was more pronounced in the presence of DMPG. The interaction of SO-6 with DMPG vesicles is stronger in the liquid-crystalline phase than in the gel phase. Extrinsic fluorescence studies indicated that the interaction of the protein with DMPG vesicles does not modify the phase transition temperature of the lipid, but decreases the amplitude of the change of fluorescence anisotropy associated with the co-operative melting of 1,6-diphenyl-1,3,5-hexatriene (DPH)-labelled vesicles. These results indicate that both electrostatic and hydrophobic components are involved in the SO-6-lipid vesicle interaction.
Hao
Q
Q
Department of Molecular Biology, Jilin University, Changchun, China.
Ding
T
T
Zhang
Y
Y
Liu
G
G
Yan
L
L
Gao
G
G
Yan
G
G
Yao
Q
Q
Li
Q
Q
D020106:Acrylamide; D000178:Acrylamides / Q000737:chemistry; D055598:Chemical Phenomena; D002627:Chemistry, Physical; D004134:Dimyristoylphosphatidylcholine / Q000737:chemistry; D005454:Fluorescence Polarization; D000922:Immunotoxins; D009699:N-Glycosyl Hydrolases; D010715:Phosphatidylglycerols / Q000737:chemistry; D010743:Phospholipids / Q000737:chemistry*; D010940:Plant Proteins / Q000737:chemistry*; D054789:Ribosome Inactivating Proteins, Type 1; D000076985:Saporins; D013050:Spectrometry, Fluorescence; D014364:Tryptophan / Q000737:chemistry
D000178:Acrylamides; D000922:Immunotoxins; D010715:Phosphatidylglycerols; D010743:Phospholipids; D010940:Plant Proteins; D054789:Ribosome Inactivating Proteins, Type 1; D020106:Acrylamide; D014364:Tryptophan; C002773:dimyristoylphosphatidylglycerol; D009699:N-Glycosyl Hydrolases; D000076985:Saporins; D004134:Dimyristoylphosphatidylcholine