Novel and highly sensitive fluorescent assay for leucine aminopeptidases

被引：48

作者：

Huang, Huazhang

Tanaka, Hiromasa

Hammock, Bruce D.

Morisseau, Christophe ^{[1
]}

机构：

[1] Univ Calif Davis, Dept Entomol, Davis, CA 95616 USA

来源：

ANALYTICAL BIOCHEMISTRY | 2009年 / 391卷 / 01期

基金：

美国国家卫生研究院;

关键词：

Metallopeptidases; N-terminal hydrolysis; Fluorescent assay; HTS assay; ENDOMETRIAL CARCINOMA; ELECTRONIC-SPECTRA; DESIGN; INVOLVEMENT; INHIBITORS; OXYTOCIN; VITRO; VIVO; LAP;

D O I：

10.1016/j.ab.2009.05.004

中图分类号：

Q5 [生物化学];

学科分类号：

071010 ; 081704 ;

摘要：

l-Leucine aminopeptidases (LAPs) are implicated in the progress of many pathological disorders and play some regulatory roles in tumor cell proliferation, invasion, and/or angiogenesis. Thus, LAPs not only could become new diagnostic or prognostic biomarkers but also may have potential as novel molecular targets for the treatment of several cancers. Highly sensitive assays are critical for early detection of changes in LAP activity and for screening potent LAP inhibitors. In this Study, we developed a novel and highly sensitive fluorescent assay for LAPs based on Substituted aminopyridines as fluorescent reporters. This assay was at least 100- and 20-fold more sensitive than commercial colorimetric and fluorescent LAP substrates, respectively. We also showed that this assay was a useful tool for monitoring LAP activities in extracts from cancer cell lines, as well as for the high-throughput screening of inhibitors, which could lead to new cancer treatments. (C) 2009 Elsevier Inc. All rights reserved.

引用

页码：11 / 16

页数：6

共 32 条

[1] DUAL FLUORESCENCE OF 2-(4'-(N,N-DIMETHYLAMINO)PHENYL)BENZOTHIAZOLE AND ITS BENZIMIDAZOLE ANALOG - EFFECT OF SOLVENT AND PH ON ELECTRONIC-SPECTRA [J].