A high throughput assay to identify small molecule modulators of prostatic acid phosphatase.
A high throughput assay to identify small molecule modulators of prostatic acid phosphatase.
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DOI：
10.2174/1875397300903010042
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发表时间：
2009-06-16
期刊：
影响因子：
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通讯作者：
Scott JE
中科院分区：
文献类型：
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作者：
Larsen RS;Zylka MJ;Scott JE
前列腺酸性磷酸酶（PAP）在伤害感受神经元中表达，并作为外核苷酸酶发挥作用。注射PAP的分泌亚型在慢性疼痛的小鼠模型中具有有效的抗伤害感受作用。这些数据表明，PAP的小分子活化剂可能具有作为慢性疼痛的新型治疗剂的效用，而抑制剂可用于在体外和体内急性抑制PAP。为了鉴定PAP活性的小分子调节剂，我们验证了高通量的基于荧光的生物化学测定，然后使用该测定来筛选化合物文库。我们通过从最终测定荧光中减去化合物荧光来降低假阳性激活剂的频率。这种方法显著减少了筛选中发现的假阳性激活剂的数量。虽然没有激活剂被证实，七个新的PAP抑制剂被确定。我们的研究结果表明，这种高通量测定可用于鉴定PAP活性的小分子调节剂。
Prostatic acid phosphatase (PAP) is expressed in nociceptive neurons and functions as an ectonucleotidase. Injection of the secretory isoform of PAP has potent antinociceptive effects in mouse models of chronic pain. These data suggested that a small molecule activator of PAP may have utility as a novel therapeutic for chronic pain, while inhibitors could be used to acutely inhibit PAP in vitro and in vivo. To identify small molecule modulators of PAP activity, we validated a high throughput, fluorescence-based biochemical assay and then used this assay to screen a compound library. We decreased the frequency of false positive activators by subtracting compound fluorescence from the final assay fluorescence. This approach significantly reduced the number of false positive activators found in the screen. While no activators were confirmed, seven novel inhibitors of PAP were identified. Our results suggest this high throughput assay could be used to identify small molecule modulators of PAP activity.