Cytotoxic effects of dynorphins through nonopioid intracellular mechanisms.
Dynorphin A, a prodynorphin-derived peptide, is able to induce neurological dysfunction and neuronal death. To study dynorphin cytotoxicity in vitro, prodynorphin-derived peptides were added into the culture medium of nonneuronal and neuronal cells or delivered into these cells by lipofection or electroporation. Cells were unaffected by extracellular exposure when peptides were added to the medium. In contrast, the number of viable cells was significantly reduced when dynorphin A or "big dynorphin," consisting of dynorphins A and B, was transfected into cells. Big dynorphin was more potent than dynorphin A, whereas dynorphin B; dynorphin B-29; [Arg(11,13)]-dynorphin A(-13)-Gly-NH-(CH(2))(5)-NH(2), a selective kappa-opioid receptor agonist; and poly-l-lysine, a basic peptide more positively charged than big dynorphin, failed to affect cell viability. The opioid antagonist naloxone did not prevent big dynorphin cytotoxicity. Thus, the toxic effects were structure selective but not mediated through opioid receptors. When big dynorphin was delivered into cells by lipofection, it became localized predominantly in the cytoplasm and not in the nuclei. Big dynorphin appeared to induce toxicity through an apoptotic mechanism that may involve synergistic interactions with the p53 tumor-suppressor protein. It is proposed that big dynorphin induces cell death by virtue of its net positive charge and clusters of basic amino acids that mimic (and thereby perhaps interfere with) basic domains involved in protein-protein interactions. These effects may be relevant for a pathophysiological role of dynorphins in the brain and spinal cord and for control of death of tumor cells, which express prodynorphin at high levels.
Tan-No
K
K
Section of Alcohol and Drug Addiction Research, Karolinska Institute, S-171 76 Stockholm, Sweden.
Cebers
G
G
Yakovleva
T
T
Hoon Goh
B
B
Gileva
I
I
Reznikov
K
K
Aguilar-Santelises
M
M
Hauser
K F
KF
Terenius
L
L
Bakalkin
G
G
D017209:Apoptosis / Q000187:drug effects / Q000502:physiology*; D002411:Cation Exchange Resins / Q000493:pharmacokinetics; D002451:Cell Compartmentation / Q000502:physiology; D002470:Cell Survival / Q000187:drug effects / Q000502:physiology; D002490:Central Nervous System / Q000187:drug effects / Q000378:metabolism / Q000503:physiopathology; D003593:Cytoplasm / Q000187:drug effects / Q000378:metabolism; D003603:Cytotoxins / Q000494:pharmacology*; D004399:Dynorphins / Q000378:metabolism / Q000633:toxicity*; D004745:Enkephalins / Q000378:metabolism; D007150:Immunohistochemistry; D008055:Lipids / Q000493:pharmacokinetics; D009270:Naloxone / Q000494:pharmacology; D009292:Narcotic Antagonists / Q000494:pharmacology; D009410:Nerve Degeneration / Q000139:chemically induced / Q000378:metabolism* / Q000503:physiopathology; D010446:Peptide Fragments / Q000494:pharmacology*; D011498:Protein Precursors / Q000378:metabolism; D017434:Protein Structure, Tertiary / Q000502:physiology; D011957:Receptors, Opioid / Q000378:metabolism*; D017473:Receptors, Opioid, kappa / Q000819:agonists / Q000378:metabolism; D014158:Transcription, Genetic / Q000187:drug effects / Q000502:physiology; D014407:Tumor Cells, Cultured / Q000166:cytology / Q000187:drug effects / Q000378:metabolism; D016159:Tumor Suppressor Protein p53 / Q000187:drug effects / Q000378:metabolism*
D002411:Cation Exchange Resins; D003603:Cytotoxins; D004745:Enkephalins; D008055:Lipids; C086724:Lipofectamine; D009292:Narcotic Antagonists; D010446:Peptide Fragments; D011498:Protein Precursors; D011957:Receptors, Opioid; D017473:Receptors, Opioid, kappa; D016159:Tumor Suppressor Protein p53; D009270:Naloxone; D004399:Dynorphins; C033660:preproenkephalin