Ectodomain shedding of CD99 within highly conserved regions is mediated by the metalloprotease meprin β and promotes transendothelial cell migration
Ectodomain shedding of CD99 within highly conserved regions is mediated by the metalloprotease meprin β and promotes transendothelial cell migration
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DOI：
10.1096/fj.201601113r
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发表时间：
2017-03-01
期刊：
影响因子：
4.8
通讯作者：
Becker-Pauly, Christoph
中科院分区：
文献类型：
--
作者：
Bedau, Tillmann;Peters, Florian;Becker-Pauly, Christoph
粘附分子CD 99对于白细胞的跨内皮迁移至关重要。在这项研究中，我们使用的生化和细胞分析表明，CD 99经历胞外脱落的金属蛋白酶meprin β和随后的膜内γ-分泌酶的蛋白水解。通过质谱法在不同脊椎动物物种高度保守的酸性区域内鉴定了CD 99中的切割位点。这一发现完全符合meprin β的独特切割特异性，具有对天冬氨酸残基的强烈偏好，并表明蛋白酶和底物的共同进化。我们假设，由meprin β进行的有限的CD 99切割将改变组织重塑过程中的细胞跨内皮迁移（TEM）行为，如炎症和癌症。事实上，在体外TEM测定中，meprin β诱导了刘易斯肺癌细胞的细胞迁移。因此，Mep 1b（-/-）小鼠中meprin β缺乏导致肺中CD 99蛋白水平显著增加。因此，meprin beta可以作为治疗靶点，因为在概念验证方法中，我们显示了meprin beta治疗小鼠的肺中CD 99蛋白的积累。Bedau，T.，彼得斯，F.，Prox，J.，Arnold，P.，施密特，F.，Finkernagel，M.，Kollmann，S.，维歇特河，Otte，A.，Ohler，A.，Stirnberg，M.，卢修斯河，Koudelka，T.，Tholey，A.，Biasin，V.，彼得齐克角美国、Kwapiszewska，G.，Becker- Pauly，C. CD 99在高度保守区域内的胞外结构域脱落由金属蛋白酶meprin B介导，并促进跨内皮细胞迁移。
The adhesion molecule CD99 is essential for the transendothelial migration of leukocytes. In this study, we used biochemical and cellular assays to show that CD99 undergoes ectodomain shedding by the metalloprotease meprin beta and subsequent intramembrane proteolysis by gamma-secretase. The cleavage site in CD99 was identified by mass spectrometry within an acidic region highly conserved through different vertebrate species. This finding fits perfectly to the unique cleavage specificity of meprin beta with a strong preference for aspartate residues and suggests coevolution of protease and substrate. We hypothesized that limited CD99 cleavage by meprin beta would alter cellular transendothelial migration (TEM) behavior in tissue remodeling processes, such as inflammation and cancer. Indeed, meprin beta induced cell migration of Lewis lung carcinoma cells in an in vitro TEM assay. Accordingly, deficiency of meprin beta in Mep1b(-/-) mice resulted in significantly increased CD99 protein levels in the lung. Therefore, meprin beta could serve as a therapeutic target, given that in aproof-of-concept approach we showed accumulation of CD99 protein in lungs of meprin beta inhibitortreated mice.- Bedau, T., Peters, F., Prox, J., Arnold, P., Schmidt, F., Finkernagel, M., Kollmann, S., Wichert, R., Otte, A., Ohler, A., Stirnberg, M., Lucius, R., Koudelka, T., Tholey, A., Biasin, V., Pietrzik, C. U., Kwapiszewska, G., Becker- Pauly, C. Ectodomain shedding of CD99 within highly conserved regions is mediated by the metalloprotease meprin b and promotes transendothelial cell migration.