A novel strategy using single-chain antibody to show the importance of Bcl-2 in mast cell survival.
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.
Cohen-Saidon
Cellina
C
Department of Biochemistry, Hebrew University Hadassah Medical School, Jerusalem, Israel.
Nechushtan
Hovav
H
Kahlon
Shira
S
Livni
Nadav
N
Nissim
Ahuva
A
Razin
Ehud
E
D000595:Amino Acid Sequence; D000818:Animals; D000906:Antibodies / Q000378:metabolism; D000918:Antibody Specificity; D017209:Apoptosis / Q000276:immunology; D001943:Breast Neoplasms; D002470:Cell Survival / Q000276:immunology; D003001:Cloning, Molecular; D015696:Gene Products, tat / Q000235:genetics; D006801:Humans; D015471:Leukemia, Basophilic, Acute; D008407:Mast Cells / Q000166:cytology* / Q000378:metabolism*; D008564:Membrane Potentials / Q000276:immunology; D008928:Mitochondria / Q000502:physiology; D008969:Molecular Sequence Data; D019151:Peptide Library; D017434:Protein Structure, Tertiary; D011518:Proto-Oncogene Proteins / Q000276:immunology / Q000378:metabolism; D019253:Proto-Oncogene Proteins c-bcl-2 / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism*; D051381:Rats; D014407:Tumor Cells, Cultured; D051028:bcl-2-Associated X Protein
D000906:Antibodies; D015696:Gene Products, tat; D019151:Peptide Library; D011518:Proto-Oncogene Proteins; D019253:Proto-Oncogene Proteins c-bcl-2; D051028:bcl-2-Associated X Protein