Abstract
Bax is a proapoptotic member of the Bcl-2 family. Members of this family can promote either cell survival, as in the case of Bcl-2 and Bcl-XL, or cell death, as in the case of Bax and Bak. Bax was first identified as a Bcl-2 binding partner by immunoprecipitation (1). Subsequently it was shown that overexpression of Bax can accelerate cell death in response to various apoptosis stimuli (2). Physiologically, Bax plays an important role in neuronal development and spermatogenesis. Animals that are deficient in Bax have increased number of neurons and the males are known to be sterile (3,4) Under pathological conditions such as cerebral and cardiac ischemia, upregulation of Bax has been reported in the afflicted area of the tissues, implicating the participation of this protein in promoting neuronal and cardiomyocytic cell death (5–7). In certain cases of human colorectal cancer, mutations were found in the gene encoding Bax, suggesting that inactivation of Bax promotes tumorigenesis by enabling the tumor cells to be less susceptible to cell death (8).
This is a preview of subscription content, log in via an institution to check access.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
References
Oltvai, Z. N., Milliman, C. L., and Korsmeyer, S. J. (1993) Bcl-2 hetero-dimerizes in vivo with a conserved homolog, Bax, that accelerates programmed cell death. Cell 74, 609–619.
Yang, E. and Korsmeyer, S. J. (1996) Molecular thanatopsis: a discourse on the Bcl2 family and cell death. Blood 88, 386–401.
Knudson, C. M., Tung, K. S. K., Tourtellotte, W. G., Brown, G. A. J., and Korsmeyer, S. J. (1995) Bax-deficient mice with lymphoic hyperplasia and male germ cell death. Science 270, 96–99.
Rodriguez, I., Ody, C., Araki, K., Garcia, I., and Vassalli, P. (1997) An early and massive wave of germinal cell apoptosis is required for the development of functional spermatogenesis. EMBO J. 16, 2262–2270.
Krajewski, S., Mai, J. K., Krajewska, M., Sikorska, M., Mossakowski, M. J., and Reed, J. C. (1995) Upregulation of Bax protein levels in neurons following cerebral ischemia. J. Neurosci. 15, 6364–6376.
Cheng, W., Kajstura, J., Nitahara, J. A., et al. (1996) Programmed myocyte cell death affects the viable myocardium after infarction in rats. Exp. Cell Res. 226, 316–327.
Misao, J., Hayakawa, Y., Ohno, M., Kato, S., Fujiwara, T., and Fujiwara, H. (1996) Expression of bcl-2 protein, an inhibitor of apoptosis, and Bax, an accelerator of apoptosis, in ventricular myocytes of human hearts with myocardial infarction. Circulation 94, 1506–1512.
Rampino, N., Yamamoto, H., Ionov, Y., et al. (1997) Somatic frameshift mutations in the BAX gene in colon cancers of the microsatellite mutator phenotype. Science 275, 967–969.
Yin, X.-M., Oltvai, Z. N., and Korsmeyer, S. J. (1994) BH1 and BH2 domains of Bcl-2 are required for inhibition of apoptosis and heterodimerization with Bax. Nature 369, 321–323.
Zha, H., C. Aime-Sempe, Sato, T., and Reed, J. C. (1996) Proapoptotic protein Bax heterodimerizes with Bcl-2 and homodimerizes with Bax via a novel domain (BH3) distinct from BH1 and BH2. J. Biol. Chem. 271, 7440–7444.
Hockenbery, D., Nunez, G., Milliman, C., Schreiber, R. D., and Korsmeyer, S. J. (1990) Bcl-2 is an inner mitochondrial membrane protein that blocks programmed cell death. Nature 348, 334–336.
Krajewski, S., Tanaka, S., Takayama, S., Schibler, M. J., Fenton, W., and Reed, J. C. (1993) Investigation of the subcellular distribution of the bcl-2 oncoprotein: residence in the nuclear envelope, endoplasmic reticulum, and outer mitochondrial membranes. Cancer Res. 53, 4701–4714.
Suzuki, M., Youle, R. J., and Tjandra, N. (2000) Structure of Bax. Coregulation of dimer formation and intracellular localization. Cell 103, 645–654.
Muchmore, S. W., Sattler, M., Liang, H., et al. (1996) X-ray and NMR structure of human Bcl-XL, an inhibitor of programmed cell death. Nature 381, 335–341.
Kagan, B. L., Finkelstein, A., and Colombini, M. (1981) Diphtheria toxin fragment forms large pores in phospholipid bilayer membranes. Proc. Natl. Acad. Sci. USA 78, 4950–4954.
Hu, V. W. and Holmes, R. K. (1984) Evidence for direct insertion of fragments A and B of diphtheria toxin into model membranes. J. Biol. Chem. 259, 12,226–12,233.
Zhan, H., Oh, K. J., Shin, Y.-K., Hubbell, W. L., and Collier, R. J. (1995) Interaction of the isolated transmembrane domain of diphtheria toxin with membranes. Biochemistry 34, 4856–4863.
Hsu, Y.-T., Wolter, K. G., and Youle, R. J. (1997) Cytosol-to-membrane redistribution of Bax and Bcl-XL during apoptosis. Proc. Natl. Acad. Sci. USA 3668–3672.
Hsu, Y.-T. and Youle, R. Y. (1998) Bax in murine thymus is a soluble monomeric protein that displays differential detergent-induced conformations. J. Biol. Chem. 273, 10,777–10,783.
Gross, A., Jockel, J., Wei, M. C., and Korsmeyer, S. J. (1998) Enforced dimerization of Bax results in its translocation, mitochondrial dysfunction and apoptosis. EMBO J. 17, 3878–3885.
Wolter, K. G., Hsu, Y.-T., Smith, C. L., Nechushtan, A., Xi, X.-G., and Youle, R. J. (1997) Movement of Bax from the cytosol to mitochondria during apoptosis. J. Cell Biol. 139, 1281–1292.
Saikumar, P., Dong, Z., Patel, Y., et al. (1998) Role of hypoxia-induced Bax translocation and cytochrome c release in reoxygenation injury. Oncogene 17, 3401–3415.
Zhang, H., Heim, J., and Meyhack, B. (1998) Redistribution of Bax from cytosol to membranes is induced by apoptotic stimuli and is an early step in the apoptotic pathway. Biochem. Biophys. Res. Commun. 251, 454–459.
Finucane, D. M., Bossy-Wetzel, E., Waterhouse, N. J., Cotter, T. G., and Green, D. R. (1999) Bax-induced caspase activation and apoptosis via cytochrome c release from mitochondria is inhibitable by Bcl-XL. J. Biol. Chem. 274, 2225–2233.
Khaled, A. R., Kim, K., Hofmeister, R., Muegge, K., and Durum, S. K. (1999) Withdrawal of IL-7 induces Bax translocation from cytosol to mitochondria through a rise in intracellular pH. Proc. Natl. Acad. Sci. USA 96, 14,476–14,481.
Murphy, K. M., Streips, U. N., and Lock, R. B. (1999) Bax membrane insertion during Fas(CD95)-induced apoptosis precedes cytochrome c release and is inhibited by Bcl-2. Oncogene 18, 5991–5999.
Putcha, G. V., Deshmukh, M., and Jr., E. M. J. (1999) BAX translocation is a critical event in neuronal apoptosis: regulation by neuroprotectants, BCL-2, and caspases. J. Neurosci. 19, 7476–7485.
Ghatan, S., Larner, S., Kinoshita, Y., et al. (2000) p38 MAP kinase mediates bax translocation in nitric oxide-induced apoptosis in neurons. J. Cell Biol. 150, 335–348.
Nechushtan, A., Smith, C. L., Hsu, Y.-T., and Youle, R. J. (1999) Conformation of the Bax C-terminus regulates subcellular location and cell death. EMBOJ. 18, 2330–2341.
Manon, S., Chaudhuri, B., and Guerin, M. (1997) Release of cytochrome c and decrease of cytochrome c oxidase in Bax-expressing yeast cells, and prevention of these effects by coexpression of Bcl-XL. FEBS Lett. 415, 29–32.
Jurgensmeier, J. M., Xie, Z., Deveraux, Q., Ellerby, L., Bredesen, D., and Reed, J. C. (1998) Bax directly induces release of cytochrome c from isolated mitochondria. Proc. Natl. Acad. Sci. USA 95, 4997–5002.
Pastorino, J. G., Chen, S.-T., Tafani, M., Snyder, J. W., and Farber, J. L. (1998) The overexpression of Bax produces cell death upon induction of the mitochondrial permeability transition. J. Biol. Chem. 273, 7770–7775.
Schlesinger, P. H., Gross, A., Yin, X. M., et al. (1997) Comparison of the ion channel characteristics of proapoptotic Bax and antiapoptotic Bcl-2. Proc. Natl. Acad. Sci. USA 94, 11,357–11,362.
Basanez, G., Nechushtan, A., Drozhinin, O., et al. (1999) Bax, but not Bcl-XL, decreases the lifetime of planar phospholipid bilayer membranes at subnanomolar concentrations. Proc. Natl. Acad. Sci. USA 96, 5492–5497.
Shimizu, S., Narita, M., and Tsujimoto, Y. (1999) Bcl-2 family proteins regulate the release of apoptogenic cytochrome c by the mitochondrial channel VDAC. Nature 399, 483–487.
Marzo, I., Brenner, C., Zamzami, N., et al. (1998) Bax and adenine nucleotide translocator cooperate in the mitochondrial control of apoptosis. Science 281, 2027–2031.
Li, P., Nijhawan, D., Budihardjo, I., et al. (1997) Cytochrome c and dATP-dependent formation of Apaf-1/caspase-9 complex initiates an apoptotic protease cascade. Cell 91, 479–489.
Murphy, K. M., Ranganathan, V., Farnsworth, M. L., Kavallaris, M., and Lock, R. B. (2000) Bcl-2 inhibits bax translocation from cytosol to mitochondria during drug-induced apoptosis of human tumor cells-111. Cell Death Differ. 7, 102–111.
Author information
Authors and Affiliations
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2002 Humana Press Inc., Totowa, NJ
About this protocol
Cite this protocol
Hsu, YT., Smaili, S. (2002). Molecular Characterization of the Proapoptotic Protein Bax. In: LeBlanc, A.C. (eds) Apoptosis Techniques and Protocols. Neuromethods, vol 37. Springer, Totowa, NJ. https://doi.org/10.1385/1-59259-188-4:001
Download citation
DOI: https://doi.org/10.1385/1-59259-188-4:001
Publisher Name: Springer, Totowa, NJ
Print ISBN: 978-1-58829-012-0
Online ISBN: 978-1-59259-188-6
eBook Packages: Springer Protocols