Larsen Rylan S, Zylka Mark J, Scott John E
Department of Cell and Molecular Physiology, UNC Neuroscience Center, University of North Carolina at Chapel Hill, CB#7545, Chapel Hill, NC 27599, USA.
Curr Chem Genomics. 2009 Jun 16;3:42-9. doi: 10.2174/1875397300903010042.
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.
前列腺酸性磷酸酶（PAP）在伤害性神经元中表达，并作为一种外切核苷酸酶发挥作用。注射PAP的分泌型异构体在慢性疼痛小鼠模型中具有强大的抗伤害感受作用。这些数据表明，PAP的小分子激活剂可能作为一种新型慢性疼痛治疗药物具有应用价值，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，我们验证了一种基于荧光的高通量生化检测方法，然后使用该检测方法筛选化合物库。我们通过从最终检测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数量。虽然没有确认激活剂，但鉴定出了七种新型PAP抑制剂。我们的结果表明，这种高通量检测方法可用于鉴定PAP活性的小分子调节剂。
