Receptor-mediated Uptake of an Extracellular Bcl-x(L) Fusion Protein Inhibits Apoptosis

Overview

Journal Proc Natl Acad Sci U S A

Specialty Science

Date 1999 Aug 18

PMID 10449732

Citations 6

Authors

X H Liu

J C Castelli

R J Youle

Affiliations

Soon will be listed here.

Abstract

Bcl-x(L), a member of the Bcl-2 family, inhibits many pathways of apoptosis when overexpressed in the cell cytosol. We examined the capacity of Bcl-x(L) fusion proteins to bind cells from the outside and block apoptosis. Full-length Bcl-x(L) protein at micromolar concentrations did not affect apoptosis when added to cell media. To increase uptake by cells, Bcl-x(L) was fused to the receptor-binding domain of diphtheria toxin (DTR). The Bcl-x(L)-DTR fusion protein blocked apoptosis induced by staurosporine, gamma-irradiation, and poliovirus in a variety of cell types when added to media. The potency of inhibition of poliovirus-induced apoptosis by Bcl-x(L)-DTR was greater than that of strong caspase inhibitors. Brefeldin A, an inhibitor of vesicular traffic between the endoplasmic reticulum and Golgi apparatus, prevented the Bcl-x(L)-DTR blockade of apoptosis induced by staurosporine, suggesting that Bcl-x(L)-DTR must be endocytosed and reach intracellular compartments for activity. Many diseases are caused by overexpression or underexpression of Bcl-x(L) homologues. Extracellular delivery of Bcl-2 family member proteins may have a wide range of uses in promoting or preventing cell death.

Citing Articles

Molecular dissection of botulinum neurotoxin reveals interdomain chaperone function.

Fischer A, Montal M Toxicon. 2013; 75:101-7.

PMID: 23396042 PMC: 3797153. DOI: 10.1016/j.toxicon.2013.01.007.

Convection enhanced delivery for treating brain tumors and selected neurological disorders: symposium review.

Vogelbaum M J Neurooncol. 2007; 83(1):97-109.

PMID: 17203397 DOI: 10.1007/s11060-006-9308-9.

The taming of the cell penetrating domain of the HIV Tat: myths and realities.

Chauhan A, Tikoo A, Kapur A, Singh M J Control Release. 2007; 117(2):148-62.

PMID: 17196289 PMC: 1859861. DOI: 10.1016/j.jconrel.2006.10.031.

Inhibition of mitochondrial neural cell death pathways by protein transduction of Bcl-2 family proteins.

Soane L, Fiskum G J Bioenerg Biomembr. 2005; 37(3):179-90.

PMID: 16167175 PMC: 2570496. DOI: 10.1007/s10863-005-6590-8.

Carrier-based strategies for targeting protein and peptide drugs to the lungs.

Cryan S AAPS J. 2005; 7(1):E20-41.

PMID: 16146340 PMC: 2751494. DOI: 10.1208/aapsj070104.

References

Donovan J, Simon M, Draper R, Montal M . Diphtheria toxin forms transmembrane channels in planar lipid bilayers. Proc Natl Acad Sci U S A. 1981; 78(1):172-6. PMC: 319013. DOI: 10.1073/pnas.78.1.172. View

Basanez G, Nechushtan A, Drozhinin O, Chanturiya A, Choe E, Tutt S . Bax, but not Bcl-xL, decreases the lifetime of planar phospholipid bilayer membranes at subnanomolar concentrations. Proc Natl Acad Sci U S A. 1999; 96(10):5492-7. PMC: 21887. DOI: 10.1073/pnas.96.10.5492. View

PAPPENHEIMER Jr A . Diphtheria: studies on the biology of an infectious disease. Harvey Lect. 1980; 76:45-73. View

Greenfield L, Bjorn M, Horn G, Fong D, Buck G, Collier R . Nucleotide sequence of the structural gene for diphtheria toxin carried by corynebacteriophage beta. Proc Natl Acad Sci U S A. 1983; 80(22):6853-7. PMC: 390084. DOI: 10.1073/pnas.80.22.6853. View

Greenfield L, Johnson V, Youle R . Mutations in diphtheria toxin separate binding from entry and amplify immunotoxin selectivity. Science. 1987; 238(4826):536-9. DOI: 10.1126/science.3498987. View

Stenmark H, Moskaug J, Madshus I, Sandvig K, Olsnes S . Peptides fused to the amino-terminal end of diphtheria toxin are translocated to the cytosol. J Cell Biol. 1991; 113(5):1025-32. PMC: 2289021. DOI: 10.1083/jcb.113.5.1025. View

Lippincott-Schwartz J, Yuan L, Tipper C, Amherdt M, Orci L, Klausner R . Brefeldin A's effects on endosomes, lysosomes, and the TGN suggest a general mechanism for regulating organelle structure and membrane traffic. Cell. 1991; 67(3):601-16. DOI: 10.1016/0092-8674(91)90534-6. View

Hunziker W, Whitney J, Mellman I . Selective inhibition of transcytosis by brefeldin A in MDCK cells. Cell. 1991; 67(3):617-27. DOI: 10.1016/0092-8674(91)90535-7. View

Prior T, Fitzgerald D, Pastan I . Translocation mediated by domain II of Pseudomonas exotoxin A: transport of barnase into the cytosol. Biochemistry. 1992; 31(14):3555-9. DOI: 10.1021/bi00129a001. View

10.

Krajewski S, Tanaka S, Takayama S, Schibler M, Fenton W, Reed J . Investigation of the subcellular distribution of the bcl-2 oncoprotein: residence in the nuclear envelope, endoplasmic reticulum, and outer mitochondrial membranes. Cancer Res. 1993; 53(19):4701-14. View

11.

Wiedlocha A, Falnes P, Madshus I, Sandvig K, Olsnes S . Dual mode of signal transduction by externally added acidic fibroblast growth factor. Cell. 1994; 76(6):1039-51. DOI: 10.1016/0092-8674(94)90381-6. View

12.

Martinou J, Dubois-Dauphin M, Staple J, Rodriguez I, Frankowski H, Missotten M . Overexpression of BCL-2 in transgenic mice protects neurons from naturally occurring cell death and experimental ischemia. Neuron. 1994; 13(4):1017-30. DOI: 10.1016/0896-6273(94)90266-6. View

13.

Chiu V, Walsh C, Liu C, Reed J, Clark W . Bcl-2 blocks degranulation but not fas-based cell-mediated cytotoxicity. J Immunol. 1995; 154(5):2023-32. View

14.

Zhan H, Oh K, Shin Y, Hubbell W, Collier R . Interaction of the isolated transmembrane domain of diphtheria toxin with membranes. Biochemistry. 1995; 34(14):4856-63. DOI: 10.1021/bi00014a043. View

15.

Memon S, Moreno M, Petrak D, Zacharchuk C . Bcl-2 blocks glucocorticoid- but not Fas- or activation-induced apoptosis in a T cell hybridoma. J Immunol. 1995; 155(10):4644-52. View

16.

Henkart P . ICE family proteases: mediators of all apoptotic cell death?. Immunity. 1996; 4(3):195-201. DOI: 10.1016/s1074-7613(00)80428-8. View

17.

Chinnaiyan A, Orth K, ORourke K, Duan H, Poirier G, Dixit V . Molecular ordering of the cell death pathway. Bcl-2 and Bcl-xL function upstream of the CED-3-like apoptotic proteases. J Biol Chem. 1996; 271(9):4573-6. DOI: 10.1074/jbc.271.9.4573. View

18.

Muchmore S, Sattler M, Liang H, Meadows R, Harlan J, Yoon H . X-ray and NMR structure of human Bcl-xL, an inhibitor of programmed cell death. Nature. 1996; 381(6580):335-41. DOI: 10.1038/381335a0. View

19.

Blanke S, Milne J, Benson E, Collier R . Fused polycationic peptide mediates delivery of diphtheria toxin A chain to the cytosol in the presence of anthrax protective antigen. Proc Natl Acad Sci U S A. 1996; 93(16):8437-42. PMC: 38689. DOI: 10.1073/pnas.93.16.8437. View

20.

Xiang J, Chao D, Korsmeyer S . BAX-induced cell death may not require interleukin 1 beta-converting enzyme-like proteases. Proc Natl Acad Sci U S A. 1996; 93(25):14559-63. PMC: 26172. DOI: 10.1073/pnas.93.25.14559. View