Cobra venom cardiotoxin (cytotoxin) isoforms and neurotoxin: comparative potency of protein kinase C inhibition and cancer cell cytotoxicity and modes of enzyme inhibition.
Effects of cobra cardiotoxin (cytotoxin) and its isoforms, and neurotoxin, on protein kinase C (PKC) and cancer cells were investigated. A positive correlation existed between hydrophobicities and activities of the toxins to inhibit PKC activity (assayed using phosphatidylserine vesicle, arachidonate monomer, and Triton/phosphatidylserine mixed micelle systems), phorbol ester binding to PKC, proliferation of several cancer cell lines, and phorbol ester-induced HL60 cell differentiation. Their relative hydrophobicities and activities, in a decreasing order, were cardiotoxin-1 approximately cardiotoxin-3 > cardiotoxin (a mixture of isoforms) > cardiotoxin-4 >> neurotoxin (inactive). Under the mixed micelle assay system (containing 0.3% Triton X-100, 8 mol % phosphatidylserine, 2 mol % diolein, and 200 microM CaCl2), cardiotoxin inhibited PKC competitively with respect to phosphatidylserine (apparent Ki of about 0.06 mol % or 2.5 microM), and in a mixed-type manner with respect to both diolein (apparent Ki of about 0.04 mol % or 1.7 microM) and Ca2+ (apparent Ki of about 2.9 microM). On the basis of findings that IC50 (approximately 0.3 microM) of cardiotoxin for inhibition of HL60 cell proliferation and differentiation was lower than its IC50 (9 microM) for PKC inhibition in vitro in the phosphatidylserine vesicle system and that PKC inhibition was the only known molecular mechanism of cardiotoxin, it was suggested that cardiotoxin might be highly membrane interacting and that the observed cellular effects of cardiotoxin might be mediated, in part, via PKC inhibition.
Chiou
S H
SH
Institute of Biochemical Sciences, National Taiwan University, Taipei.
Raynor
R L
RL
Zheng
B
B
Chambers
T C
TC
Kuo
J F
JF
D000595:Amino Acid Sequence; D000818:Animals; D001921:Brain / Q000201:enzymology; D002454:Cell Differentiation / Q000187:drug effects; D002470:Cell Survival / Q000187:drug effects*; D003603:Cytotoxins / Q000737:chemistry / Q000494:pharmacology*; D004546:Elapid Venoms / Q000737:chemistry / Q000494:pharmacology*; D006801:Humans; D007700:Kinetics; D015464:Leukemia, Myelogenous, Chronic, BCR-ABL Positive; D015473:Leukemia, Promyelocytic, Acute; D008969:Molecular Sequence Data; D009498:Neurotoxins / Q000737:chemistry / Q000494:pharmacology*; D015240:Phorbol 12,13-Dibutyrate / Q000378:metabolism; D011493:Protein Kinase C / Q000037:antagonists & inhibitors*; D017386:Sequence Homology, Amino Acid; D013329:Structure-Activity Relationship; D013552:Swine; D013755:Tetradecanoylphorbol Acetate / Q000494:pharmacology; D014407:Tumor Cells, Cultured
D003603:Cytotoxins; D004546:Elapid Venoms; D009498:Neurotoxins; D015240:Phorbol 12,13-Dibutyrate; D011493:Protein Kinase C; D013755:Tetradecanoylphorbol Acetate