Abstract
Cell penetrating peptides (CPPs) have been used to deliver the anti-apoptotic Bcl-xL-derived BH4 peptide to prevent injury-induced apoptosis both in vitro and in vivo. Here we demonstrate that the nuclear localization sequence (NLS) from the SV40 large T antigen has favorable properties for BH4 domain delivery to lymphocytes compared to sequences based on the HIV-1 TAT sequence. While both TAT-BH4 and NLS-BH4 protected primary human mononuclear cells from radiation-induced apoptotic cell death, TAT-BH4 caused persistent membrane damage and even cell death at the highest concentrations tested (5-10 microM) and correlated with in vivo toxicity as intravenous administration of TAT-BH4 caused rapid death. The NLS-BH4 peptide has significantly attenuated toxicity compared to TAT-BH4 and we established a dosing regimen of NLS-BH4 that conferred a significant survival advantage in a post-exposure treatment model of LD90 total body irradiation.
Publication types
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Research Support, N.I.H., Extramural
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Research Support, Non-U.S. Gov't
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Research Support, U.S. Gov't, Non-P.H.S.
MeSH terms
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Amino Acid Sequence
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Animals
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Antigens, Polyomavirus Transforming / genetics
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Antigens, Polyomavirus Transforming / metabolism
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Antigens, Polyomavirus Transforming / pharmacology
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Apoptosis / drug effects*
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Cell Survival / drug effects
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Cells, Cultured
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Humans
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Leukocytes, Mononuclear / drug effects
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Leukocytes, Mononuclear / physiology
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Male
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Mice
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Mice, Inbred C57BL
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Molecular Sequence Data
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Nuclear Localization Signals / genetics
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Nuclear Localization Signals / metabolism
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Nuclear Localization Signals / pharmacology
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Peptides / pharmacology*
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Protein Structure, Tertiary
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Whole-Body Irradiation*
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bcl-X Protein / genetics
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bcl-X Protein / metabolism
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bcl-X Protein / pharmacology
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tat Gene Products, Human Immunodeficiency Virus / genetics
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tat Gene Products, Human Immunodeficiency Virus / metabolism
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tat Gene Products, Human Immunodeficiency Virus / pharmacology
Substances
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Antigens, Polyomavirus Transforming
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NLS-BH4 protein
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Nuclear Localization Signals
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Peptides
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TAT-BH4 peptide
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bcl-X Protein
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tat Gene Products, Human Immunodeficiency Virus