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Western blot analysis of Granzyme B. A NK-92 cell lysate (Human natural killer cells derived from malignant non-Hodgkin's lymphoma donor; ATCC CRL-2407) was probed with the mouse anti-granzyme B antibody at concentrations of 0.125 µg/mL (lane 1), 0.0625 µg/mL (lane 2), and 0.03125 µg/mL (lane 3). Granzyme B is identified at ~32 kDa.
BD Pharmingen™ Purified Mouse Anti-Granzyme B
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Western blot: Please refer to http://www.bdbiosciences.com/pharmingen/protocols/Western_Blotting.shtml
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The primary mechanism by which cytotoxic T cells eliminate virally infected cells is by granule exocytosis. The release of cytotoxic granule contents by cytotoxic T lymphocytes (CTL) triggers apoptotic target cell death. CTL granules contain a poreforming protein, perforin, and a group of serine proteases called granzymes. In the classic model, perforins create holes in the target cell membrane, allowing entrance of the granzymes. Granzyme A and B are the predominant granzymes activated after CTL activation, but each act via an independent apoptotic pathway; granzyme B is activated immediately, while granzyme A acts hours later. The physiological substrates for granzyme A in the apoptotic pathway have not been identified. Studies involving mice which are deficient in both granzyme A and B suggest a model whereby the granzyme B pathway may have evolved as the major apoptotic pathway with the granzyme A pathway acting as a backup. Granzyme B has been shown to induce apoptosis and to cleave a number of substrates which are similar in specificity to those of the caspase family of proteinases. Granzyme B can cleave substrates, such as DNA-PKcs, and nuclear mitotic apparatus protein (NuMA). Furthermore, Granzyme B can also cleave substrates such as Bid and DFF45 in a caspase-independent fashion. However, further research is needed to delineate the exact role of caspases in cytotoxic T lymphocyte-induced apoptosis involving Granzyme B. Granzyme B migrates at approximately 32 kDa in SDS/PAGE. Clone 2C5/F5 recognizes human and rat granzyme B.
研发参考 (8)
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Andrade F, Roy S, Nicholson D, Thornberry N, Rosen A, Casciola-Rosen L. Granzyme B directly and efficiently cleaves several downstream caspase substrates: implications for CTL-induced apoptosis. Immunity. 1998; 8(4):451-460. (Biology). 查看参考
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Barry M, Heibein JA, Pinkoski MJ, et al. Granzyme B short-circuits the need for caspase 8 activity during granule-mediated cytotoxic T-lymphocyte killing by directly cleaving Bid. Mol Cell Biol. 2000; 20(11):3781-3794. (Biology). 查看参考
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Beresford PJ, Xia Z, Greenberg AH, Lieberman J. Granzyme A loading induces rapid cytolysis and a novel form of DNA damage independently of caspase activation. Immunity. 1999; 10(5):585-594. (Biology). 查看参考
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Pinkoski MJ, Waterhouse NJ, Heibein JA, et al. Granzyme B-mediated apoptosis proceeds predominantly through a Bcl-2-inhibitable mitochondrial pathway. J Biol Chem. 2001; 276(15):12060-12067. (Biology). 查看参考
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Rotonda J, Garcia-Calvo M, Bull HG, et al. The three-dimensional structure of human granzyme B compared to caspase-3, key mediators of cell death with cleavage specificity for aspartic acid in P1. Chem Biol. 2001; 8(4):357-368. (Biology). 查看参考
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Sharif-Askari E, Alam A, Rheaume E, et al. Direct cleavage of the human DNA fragmentation factor-45 by granzyme B induces caspase-activated DNase release and DNA fragmentation. EMBO J. 2001; 20(12):3101-3113. (Biology). 查看参考
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Shresta S, Graubert TA, Thomas DA, Raptis SZ, Ley TJ. Granzyme A initiates an alternative pathway for granule-mediated apoptosis. Immunity. 1999; 10(5):595-605. (Biology). 查看参考
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Trapani JA, Smyth MJ, Apostolidis VA, Dawson M, and Browne KA. Granule serine proteases are normal nuclear constituents of natural killer cells. J Biol Chem. 1994; 269:18359-18365. (Biology). 查看参考
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