Initiation of apoptosis by granzyme B requires direct cleavage of bid, but not direct granzyme B-mediated caspase activation.
Initiation of apoptosis by granzyme B requires direct cleavage of bid, but not direct granzyme B-mediated caspase activation.
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DOI：
10.1084/jem.192.10.1403
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发表时间：
2000-11-20
期刊：
影响因子：
--
通讯作者：
Trapani JA
中科院分区：
文献类型：
--
作者：
Sutton VR;Davis JE;Cancilla M;Johnstone RW;Ruefli AA;Sedelies K;Browne KA;Trapani JA
颗粒酶B介导的细胞凋亡的重要上游步骤仍然不确定。在此，我们表明，颗粒酶B触发线粒体凋亡途径，通过直接裂解的投标，然而，裂解的半胱氨酸蛋白酶原停滞时，线粒体破坏被阻止Bcl-2。颗粒酶B抗性Bcl-2过表达FDC-P1细胞的敏感性通过共表达野生型Bid或Bid与其caspase-8切割位点的突变而恢复，并且两种类型的Bid都被切割。然而，Bid与突变的颗粒酶B切割位点保持完整，并没有恢复细胞凋亡。Bid的突变阻止其与Bcl-2的相互作用被切割，但也未能恢复凋亡。Bcl-2过表达并不延迟颗粒酶B对Bid的快速切割（<2分钟）。这些结果清楚地将Bid切割置于线粒体Bcl-2的上游。在颗粒酶B处理的Jurkat细胞中，尽管用z-Val-Ala-Asp-fluoromethylketone或Asp-Glu-Val-Asp-fluoromethylketone使胱天蛋白酶失活，但仍发生内源性Bid裂解和线粒体膜去极化的损失。无论Bcl-2过表达，观察到半胱天冬酶原-3和-8的初始部分加工;然而，后来的加工被Bcl-2完全废除。总体而言，我们的研究结果表明，线粒体扰动申办是必要的，以实现半胱天冬酶活性和细胞死亡的致死阈值，由于颗粒酶B。
The essential upstream steps in granzyme B–mediated apoptosis remain undefined. Herein, we show that granzyme B triggers the mitochondrial apoptotic pathway through direct cleavage of Bid; however, cleavage of procaspases was stalled when mitochondrial disruption was blocked by Bcl-2. The sensitivity of granzyme B–resistant Bcl-2–overexpressing FDC-P1 cells was restored by coexpression of wild-type Bid, or Bid with a mutation of its caspase-8 cleavage site, and both types of Bid were cleaved. However, Bid with a mutated granzyme B cleavage site remained intact and did not restore apoptosis. Bid with a mutation preventing its interaction with Bcl-2 was cleaved but also failed to restore apoptosis. Rapid Bid cleavage by granzyme B (<2 min) was not delayed by Bcl-2 overexpression. These results clearly placed Bid cleavage upstream of mitochondrial Bcl-2. In granzyme B–treated Jurkat cells, endogenous Bid cleavage and loss of mitochondrial membrane depolarization occurred despite caspase inactivation with z-Val-Ala-Asp-fluoromethylketone or Asp-Glu-Val-Asp-fluoromethylketone. Initial partial processing of procaspase-3 and -8 was observed irrespective of Bcl-2 overexpression; however, later processing was completely abolished by Bcl-2. Overall, our results indicate that mitochondrial perturbation by Bid is necessary to achieve a lethal threshold of caspase activity and cell death due to granzyme B.
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DOI：
10.1084/jem.189.1.131
发表时间：
1999-01-04
期刊：
The Journal of experimental medicine
影响因子：
--
作者：
MacDonald G;Shi L;Vande Velde C;Lieberman J;Greenberg AH
通讯作者：
Greenberg AH
影响因子：
64.5
作者：
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Wang, XD
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4.8
作者：
Jans, DA;Jans, P;Trapani, JA
通讯作者：
Trapani, JA
影响因子：
64.8
作者：
KAGI, D;LEDERMANN, B;HENGARTNER, H
通讯作者：
HENGARTNER, H
DOI：
10.1084/jem.177.1.195
发表时间：
1993-01-01
期刊：
The Journal of experimental medicine
影响因子：
--
作者：
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通讯作者：
Golstein P