Development of near-infrared fluorophore (NIRF)-labeled activity-based probes for in vivo imaging of legumain.
Development of near-infrared fluorophore (NIRF)-labeled activity-based probes for in vivo imaging of legumain.
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DOI：
10.1021/cb900232a
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发表时间：
2010-02-19
影响因子：
4
通讯作者：
Bogyo M
中科院分区：
文献类型：
--
作者：
Lee J;Bogyo M
天冬酰胺基内肽酶或豆荚蛋白是一种溶酶体半胱氨酸蛋白酶，最初在植物中鉴定，后来发现参与高等真核生物中的抗原呈递。Legumain在许多人类癌症中也被上调，最近的研究表明它可能在肿瘤发生过程中发挥重要的功能作用。然而，由于缺乏合适的选择性小分子抑制剂和成像试剂，在人类疾病的相关动物模型中的详细功能研究受到阻碍。在这里，我们提出的设计，优化和在体内应用的豆荚蛋白的荧光标记的活性为基础的探针（ABPs）。我们证明了优化的氮杂肽基天冬酰胺环氧化物是高度选择性和有效的抑制剂，可以很容易地转化为NIRF标记的ABP全身，非侵入性成像应用。我们表明，这些探针特异性标记豆荚蛋白在各种正常组织以及在实体瘤中应用时，在体内。有趣的是，向探针中加入细胞穿透肽增强了细胞摄取，但由于其在溶酶体中的积累，导致对其他溶酶体蛋白酶的交叉反应性增加。总之，我们发现氮杂肽基天冬酰胺ABPs是未来研究更复杂的人类疾病模型中豆荚蛋白功能的有价值的新工具。
Asparaginyl endopeptidase, or legumain, is a lysosomal cysteine protease that was originally identified in plants and later found to be involved in antigen presentation in higher eukaryotes. Legumain is also up-regulated in a number of human cancers and recent studies suggest that it may play important functional roles in the process of tumorigenesis. However, detailed functional studies in relevant animal models of human disease have been hindered by the lack of suitably selective small molecule inhibitors and imaging reagents. Here we present the design, optimization and in vivo application of fluorescently labeled activity based probes (ABPs) for legumain. We demonstrate that optimized aza-peptidyl Asn epoxides are highly selective and potent inhibitors that can be readily converted into NIRF labeled ABPs for whole body, non-invasive imaging applications. We show that these probes specifically label legumain in various normal tissues as well as in solid tumors when applied in vivo. Interestingly, addition of cell penetrating peptides to the probes enhanced cellular uptake but resulted in increased cross-reactivity towards other lysosomal proteases as the result of their accumulation in lysosomes. Overall, we find that aza-peptidyl Asn ABPs are valuable new tools for the future study of legumain function in more complex models of human disease.
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影响因子：
7.3
作者：
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通讯作者：
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影响因子：
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作者：
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通讯作者：
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作者：
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DOI：
10.1073/pnas.0809824105
发表时间：
2009-01-13
影响因子：
11.1
作者：
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通讯作者：
Ye, Keqiang