NK cell protease granzyme M targets α-tubulin and disorganizes the microtubule network

被引:61
作者
Bovenschen, Niels [1 ]
de Koning, Pieter J. A. [1 ]
Quadir, Razi [1 ]
Broekhuizen, Roel [1 ]
Damen, J. Mirjam A. [2 ]
Froelich, Christopher J. [3 ]
Slijper, Monique [2 ]
Kummer, J. Alain [1 ]
机构
[1] Univ Med Ctr, Dept Pathol, NL-3584 CX Utrecht, Netherlands
[2] Univ Utrecht, Dept Biomol Mass Spectrometry, Utrecht, Netherlands
[3] Evanston NW Healthcare Res Inst, Dept Med, Evanston, IL 60201 USA
关键词
D O I
10.4049/jimmunol.180.12.8184
中图分类号
R392 [医学免疫学]; Q939.91 [免疫学];
学科分类号
100102 ;
摘要
Serine protease granzyme M (GrM) is highly expressed in the cytolytic granules of NK cells, which eliminate virus-infected cells and tumor cells. The molecular mechanisms by which GrM induces cell death, however, remain poorly understood. In this study we used a proteomic approach to scan the native proteome of human tumor cells for intracellular substrates of GrM. Among other findings, this approach revealed several components of the cytoskeleton. GrM directly and efficiently cleaved the actin-plasma membrane linker ezrin and the microtubule component alpha-tubulin by using purified proteins, tumor cell lysates, and tumor cells undergoing cell death induced by perforin and GrM. These cleavage events occurred independently of caspases or other cysteine proteases. Kinetically, alpha-tubulin was more efficiently cleaved by GrM as compared with ezrin. Direct alpha-tubulin proteolysis by GrM is complex and occurs at multiple cleavage sites, one of them being Leu at position 269. GrM disturbed tubulin polymerization dynamics in vitro and induced microtubule network disorganization in tumor cells in vivo. We conclude that GrM targets major components of the cytoskeleton that likely contribute to NK cell-induced cell death.
引用
收藏
页码:8184 / 8191
页数:8
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