Abstract
Apoptosis or programmed cell death plays an important role in a wide variety of physiologic processes and is regulated by proteins of the Bcl-2 family consisting of both antiapoptotic and proapoptotic factors. The direct involvement of the Bcl-2 protein family in the process of mast cell apoptosis has not been clarified. In the present work we have used a single-chain antibody (scFv) raised against Bcl-2 derived from a semisynthetic human phage-display antibody library. The addition of TAT sequence, which is responsible for translocation through the membrane, endows the anti-Bcl-2-scFv with the ability to penetrate living cells. Moreover, it specifically neutralizes Bcl-2 intracellularly by binding to the BH1 domain and eradicates its antiapoptotic activity in 2 types of mast cells and in a human breast cancer cell line.
Publication types
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Research Support, Non-U.S. Gov't
MeSH terms
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Amino Acid Sequence
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Animals
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Antibodies / metabolism
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Antibody Specificity
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Apoptosis / immunology
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Breast Neoplasms
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Cell Survival / immunology
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Cloning, Molecular
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Gene Products, tat / genetics
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Humans
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Leukemia, Basophilic, Acute
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Mast Cells / cytology*
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Mast Cells / metabolism*
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Membrane Potentials / immunology
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Mitochondria / physiology
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Molecular Sequence Data
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Peptide Library
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Protein Structure, Tertiary
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Proto-Oncogene Proteins / immunology
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Proto-Oncogene Proteins / metabolism
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Proto-Oncogene Proteins c-bcl-2 / chemistry
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Proto-Oncogene Proteins c-bcl-2 / genetics*
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Proto-Oncogene Proteins c-bcl-2 / metabolism*
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Rats
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Tumor Cells, Cultured
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bcl-2-Associated X Protein
Substances
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Antibodies
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Gene Products, tat
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Peptide Library
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Proto-Oncogene Proteins
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Proto-Oncogene Proteins c-bcl-2
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bcl-2-Associated X Protein