Caissarolysin I (Bcs I), a new hemolytic toxin from the Brazilian sea anemone Bunodosoma caissarum: purification and biological characterization.
Two cationic proteins, C1 and C3, were purified to homogeneity from the hemolytic fraction of the venom of Bunodosoma caissarum sea anemone. The purification processes employed gel filtration followed by ion exchange chromatography, being the purity and molecular mass confirmed by SDS-PAGE and mass spectrometry. Protein C1 represented the second major peak of the hemolytic fraction and was previously believed to be a cytolysin belonging to a new class of hemolysins. The C1 protein has a molecular mass of 15495 Da and was assayed for hemolysis, PLA2 activity and acute toxicity in crabs and mice, showing no activity in these assays. It has an amino terminal with no similarity to all known hemolysins and, therefore, should not be considered a toxin, being its function completely unknown. The protein C3 (19757 Da), that also lacks PLA2 activity, was recognized by antiserum against Eqt II and presented high hemolytic activity to human erythrocytes (ED50 of 0.270 microg/ml), being named Caissarolysin I (Bcs I). Its activity was inhibited by pre-incubation with sphingomyelin (SM) and also when in presence of erythrocytes pre-treated with the SMase P2, a phospholipase D from the brown spider Loxosceles intermedia, indicating that SM is the main target of Bcs I. Caissarolysin I is the first hemolysin purified from a sea anemone belonging to the genus Bunodosoma and belongs to the Actinoporin family of sea anemone hemolysins.
de Oliveira
Joacir Stolarz
JS
Laboratório de Imunoquímica, Instituto Butantan, Av. Prof. Vital Brazil, 1500, CEP 05508-900, São Paulo, SP, Brazil. jstolarz@usp.br
Zaharenko
André Junqueira
AJ
de Freitas
José Carlos
JC
Konno
Katsuhiro
K
de Andrade
Sonia A
SA
Portaro
Fernanda C V
FC
Richardson
Michael
M
Sant'anna
Osvaldo Augusto
OA
Tambourgi
Denise V
DV
D000595:Amino Acid Sequence; D000818:Animals; D001681:Biological Assay; D002850:Chromatography, Gel; D002852:Chromatography, Ion Exchange; D003064:Cnidarian Venoms / Q000302:isolation & purification* / Q000378:metabolism; D004355:Drug Stability; D006460:Hemolysin Proteins / Q000187:drug effects / Q000302:isolation & purification; D006461:Hemolysis / Q000187:drug effects; D006358:Hot Temperature; D006801:Humans; D051379:Mice; D008969:Molecular Sequence Data; D008970:Molecular Weight; D010741:Phospholipases A / Q000378:metabolism; D054467:Phospholipases A2; D010727:Phosphoric Diester Hydrolases / Q000378:metabolism; D012615:Sea Anemones / Q000737:chemistry*; D013109:Sphingomyelins / Q000494:pharmacology
D003064:Cnidarian Venoms; D006460:Hemolysin Proteins; D013109:Sphingomyelins; C510096:caissarolysin I, Bunodosoma caissarum; D010741:Phospholipases A; D054467:Phospholipases A2; D010727:Phosphoric Diester Hydrolases; C055160:sphingomyelin phosphodiesterase D