Selective imaging of cathepsin L in breast cancer by fluorescent activity-based probes.
Selective imaging of cathepsin L in breast cancer by fluorescent activity-based probes.
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DOI：
10.1039/c7sc04303a
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发表时间：
2018-02-28
期刊：
影响因子：
8.4
通讯作者：
Drag M
中科院分区：
文献类型：
--
作者：
Poreba M;Rut W;Vizovisek M;Groborz K;Kasperkiewicz P;Finlay D;Vuori K;Turk D;Turk B;Salvesen GS;Drag M
高选择性荧光底物和活性探针用于监测乳腺癌细胞系组织蛋白酶L的活性。半胱氨酸组织蛋白通常在溶酶体降解系统中发挥作用，在该系统中，半胱氨酸组织蛋白对维持细胞内稳态和MHC II免疫反应至关重要，并已被发现在几种疾病和肿瘤进展中发挥重要作用。在一个复杂的蛋白质组中选择性地观察个体的蛋白水解酶活性对于确定它们在正常细胞和肿瘤细胞中的作用是非常重要的，从而有助于我们理解蛋白质分解网络的调节。一种被普遍接受的监测蛋白酶活性的方法是使用小分子底物和基于活性的探针。然而，人类有11种半胱氨酸组织蛋白，其中一些显示出重叠的底物特异性，这使得选择性靶向单一组织蛋白的小分子的开发非常具有挑战性。在这里，我们利用HyCoSuL，一种定位扫描底物的方法，开发了一种高选择性的荧光底物和基于活性的探针来监测乳腺癌细胞株MDAMB-231中组织蛋白酶L的活性。使用该探针使我们能够将组织蛋白酶L的活性与其他组织蛋白酶区分开来，特别是组织蛋白酶B，它在正常和转化的细胞类型中广泛表达。我们发现，组织蛋白酶L在MDAMB-231细胞中的定位与组织蛋白酶B的定位有很大的重叠，然而，一些富含组织蛋白酶L的溶酶体缺乏组织蛋白酶B的活性。总体而言，这些研究表明，HyCoSuL衍生的小分子探针是成像活细胞中组织蛋白酶L活性的有价值的工具。因此，这种方法能够评估组织蛋白酶L在肿瘤发生中的功能，并适用于其他半胱氨酸组织蛋白酶。
Highly-selective fluorogenic substrate and activity-based probe for monitoring cathepsin L activity in the breast cancer cell line MDA-MB-231. Cysteine cathepsins normally function in the lysosomal degradation system where they are critical for the maintenance of cellular homeostasis and the MHC II immune response, and have been found to have major roles in several diseases and in tumor progression. Selective visualization of individual protease activity within a complex proteome is of major importance to establish their roles in both normal and tumor cells, thereby facilitating our understanding of the regulation of proteolytic networks. A generally accepted means to monitor protease activity is the use of small molecule substrates and activity-based probes. However, there are eleven human cysteine cathepsins, with a few of them displaying overlapping substrate specificity, making the development of small molecules that selectively target a single cathepsin very challenging. Here, we utilized HyCoSuL, a positional scanning substrate approach, to develop a highly-selective fluorogenic substrate and activity-based probe for monitoring cathepsin L activity in the breast cancer cell line MDA-MB-231. Use of this probe enabled us to distinguish the activity of cathepsin L from that of other cathepsins, particularly cathepsin B, which is abundant and ubiquitously expressed in normal and transformed cell types. We found that cathepsin L localization in MDA-MB-231 cells greatly overlaps with that of cathepsin B, however, several cathepsin L-rich lysosomes lacked cathepsin B activity. Overall, these studies demonstrate that HyCoSuL-derived small molecule probes are valuable tools to image cathepsin L activity in living cells. This approach thus enables evaluation of cathepsin L function in tumorigenesis and is applicable to other cysteine cathepsins.
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影响因子：
4.8
作者：
Ménard, R;Carmona, E;Mort, JS
通讯作者：
Mort, JS
影响因子：
3.7
作者：
Creasy, Blaine M.;Hartmann, Constance B.;McCoy, Kathleen L.
通讯作者：
McCoy, Kathleen L.
DOI：
10.1007/978-1-4939-6439-0_11
发表时间：
2017-01-01
期刊：
ACTIVITY-BASED PROTEOMICS: METHODS AND PROTOCOLS
影响因子：
--
作者：
Edgington-Mitchell, Laura E.;Bogyo, Matthew;Verdoes, Martijn
通讯作者：
Verdoes, Martijn
影响因子：
--
作者：
Bogyo, M;Verhelst, S;Greenbaum, D
通讯作者：
Greenbaum, D
影响因子：
14.8
作者：
Blum, Galia;von Degenfeld, Georges;Bogyo, Matthew
通讯作者：
Bogyo, Matthew