A simple assay for hyaluronidase activity using fluorescence polarization
A simple assay for hyaluronidase activity using fluorescence polarization
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DOI：
10.1016/j.bbrc.2008.09.047
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发表时间：
2008-11-21
影响因子：
3.1
通讯作者：
Kawashima, Hiroto
中科院分区：
文献类型：
--
作者：
Murai, Toshiyuki;Kawashima, Hiroto
透明质酸是一种大型糖胺聚糖，是细胞外基质的主要成分，与细胞表面受体（如CD44）相互作用。我们之前报道过碎片状透明质酸，其大小在癌症患者中经常检测到，诱导CD44切割并同时增强肿瘤细胞迁移。虽然透明质酸降解为更小的片段已被发现是调节癌症进展的关键反应，但持续检测透明质酸酶活性的简单方法尚未建立。在这里，我们展示了荧光标记的透明质酸可以作为透明质酸酶活性连续测定的底物。建立了一种非常简单的测定透明质酸基油荧光偏振降解的方法。该方法将为连续测量细胞透明质酸酶活性和透明质酸与其受体的结合提供一种方法，因此对研究透明质酸在癌症进展中的作用是有用的。(C) 2008爱思唯尔公司版权所有
Hyaluronan is a large glycosaminoglycan and is a major constituent of the extracellular matrix, interacting with cell surface receptors such as CD44. We previously reported that fragmented hyaluronan, with the size frequently detected in cancer patients, induces CD44 cleavage and concomitantly enhances tumor cell migration. Although hyaluronan degradation to smaller fragments has been revealed to be a key reaction in regulating cancer progression, simple methods for continuously detecting hyaluronidase activity have not been established. Here, we show that fluorescently-labeled hyaluronan serves as a substrate for continuous assay Of hyaluronidase activity. A very simple assay was established to measure degradation of hyaluronan based oil fluorescence polarization. The developed assay method would provide a way for continuous measurement of cellular hyaluronidase activity and also for measurement of binding of hyaluronan to its receptors, and thus Should be useful for investigation of the function of hyaluronan in cancer progression. (C) 2008 Elsevier Inc. All rights reserved