Properties of St I and St II, two isotoxins isolated from Stichodactyla helianthus: a comparison.
Sticholysins I and II are two highly hemolytic polypeptides purified from the Caribbean Sea anemone Stichodactyla helianthus. Their high sequence homology (93%) indicates that they correspond to isoforms of the same hemolysin. The spectroscopic measurements show a close similarity in the secondary structure content, conformation and stability of both toxins. Exposure of the toxins to high pHs (>11), a free radical source (AAPH), urea or temperature produce permanent changes in the toxin that lead to a significant loss of HA. It is significant to note that this loss of hemolytic activity occurs when other indicators, probably with the only exception of near-UV CD spectra, barely detect changes in the protein structure. This emphasizes the sensitivity of the protein function to changes in the macromolecule conformation. The most noticeable difference between both toxins is the considerably higher activity of St II, both measured in terms of erythrocyte internal K(+) exit or hemolysis; which is related to enthalpic factors. This difference is not due to an incomplete association of St I to the membrane. We consider then that the different pore forming capacity of both toxins in erythrocytes can be explained in terms of the difference in charge of the N-terminal fragment, than can considerably reduce the St I insertion rate in the membrane probably due to the negatively charged outer leaflet of the red blood cell, without a significant reduction of its capacity to bind to the cell membrane. This electrostatic effect, together with a slightly more relaxed structure in St II, could explain the higher pore forming capacity of St II in the red blood cell membrane.
Martinez
D
D
Faculty of Biology, University of La Habana, Cuba.
Campos
A M
AM
Pazos
F
F
Alvarez
C
C
Lanio
M E
ME
Casallanovo
F
F
Schreier
S
S
Salinas
R K
RK
Vergara
C
C
Lissi
E
E
D000578:Amidines / Q000378:metabolism*; D000818:Animals; D002942:Circular Dichroism; D003064:Cnidarian Venoms / Q000737:chemistry* / Q000633:toxicity; D004912:Erythrocytes / Q000187:drug effects / Q000502:physiology*; D006460:Hemolysin Proteins / Q000187:drug effects / Q000378:metabolism*; D006801:Humans; D006863:Hydrogen-Ion Concentration; D008081:Liposomes / Q000737:chemistry* / Q000378:metabolism*; D009498:Neurotoxins / Q000737:chemistry* / Q000378:metabolism*; D016877:Oxidants / Q000378:metabolism; D011188:Potassium / Q000032:analysis / Q000502:physiology*; D011487:Protein Conformation; D020033:Protein Isoforms / Q000737:chemistry* / Q000378:metabolism*; D017433:Protein Structure, Secondary; D012615:Sea Anemones / Q000472:pathogenicity*; D017386:Sequence Homology, Amino Acid; D013050:Spectrometry, Fluorescence; D013696:Temperature; D013997:Time Factors
D000578:Amidines; D003064:Cnidarian Venoms; D006460:Hemolysin Proteins; D008081:Liposomes; D009498:Neurotoxins; D016877:Oxidants; D020033:Protein Isoforms; C059713:neurotoxin I, Stichodactyla helianthus; C046728:2,2'-azobis(2-amidinopropane); D011188:Potassium