Effects of sodium dodecyl sulfate on the conformation and hemolytic activity of St I and St II, two isotoxins purified from Stichodactyla helianthus.
The effect of sodium dodecyl sulfate (SDS) upon the conformation and hemolytic activity of St I and St II strongly depends on its concentration. At relatively low surfactant concentrations (ca. 0.5-5mM range) the surfactant leads to the formation of aggregates, as suggested by the turbidity observed even at relatively low (micromolar range) protein concentrations. In this surfactant range, the proteins show an increase in intrinsic fluorescence intensity and reduced quenching by acrylamide, with an almost total loss of its hemolytic activity. At higher surfactant concentrations the protein adducts disaggregates. This produces a decrease in fluorescence intensity, increase in quenching efficiency by acrylamide, loss of the native tertiary conformation (as reported by the near UV-CD spectra), and increase in alpha-helix content (as evidenced by the far UV-CD spectra). However, and in spite of these substantial changes, the toxins partially recover their hemolytic activity. The reasons for this recovering of the activity at high surfactant concentrations is discussed.
Lanio
M E
ME
Faculty of Biology, University of Habana, La Habana, Cuba.
Alvarez
C
C
Pazos
F
F
Martinez
D
D
Martínez
Y
Y
Casallanovo
F
F
Abuin
E
E
Schreier
S
S
Lissi
E
E
D000818:Animals; D002942:Circular Dichroism; D003064:Cnidarian Venoms / Q000737:chemistry / Q000494:pharmacology*; D004305:Dose-Response Relationship, Drug; D004912:Erythrocytes / Q000187:drug effects; D006460:Hemolysin Proteins / Q000737:chemistry / Q000494:pharmacology*; D006461:Hemolysis / Q000187:drug effects*; D006801:Humans; D009930:Organic Chemicals; D011487:Protein Conformation / Q000187:drug effects; D012615:Sea Anemones; D012967:Sodium Dodecyl Sulfate / Q000008:administration & dosage / Q000494:pharmacology*; D013050:Spectrometry, Fluorescence; D013501:Surface-Active Agents / Q000008:administration & dosage / Q000494:pharmacology*
D003064:Cnidarian Venoms; D006460:Hemolysin Proteins; D009930:Organic Chemicals; D013501:Surface-Active Agents; C404794:sticholysin II; C121348:stycholysin I; D012967:Sodium Dodecyl Sulfate