Recombinant mouse PAP has pH-dependent ectonucleotidase activity and acts through A(1)-adenosine receptors to mediate antinociception.
Recombinant mouse PAP has pH-dependent ectonucleotidase activity and acts through A(1)-adenosine receptors to mediate antinociception.
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DOI：
10.1371/journal.pone.0004248
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发表时间：
2009
期刊：
影响因子：
3.7
通讯作者：
Zylka, Mark J.
中科院分区：
文献类型：
--
作者：
Sowa, Nathaniel A.;Vadakkan, Kunjumon I.;Zylka, Mark J.
前列腺酸性磷酸酶（PAP）在痛觉神经元中表达，是一种外核苷酶。当棘内注射时，人类和牛PAP蛋白的分泌异构体具有强效和持久的抗伤害作用，这种作用依赖于a1 -腺苷受体（A1R）的激活。在这项研究中，我们使用杆状病毒表达系统纯化了小鼠(m)PAP的分泌异构体，以确定重组mPAP是否也具有抗感染性。我们发现mPAP在中性pH （pH 7.0）下可以使AMP去磷酸化，并在较小程度上使ADP去磷酸化。相反，mPAP在酸性pH （pH 5.6）下使所有嘌呤核苷酸（AMP， ADP， ATP）去磷酸化。mPAP的跨膜异构体具有类似的ph依赖性外核苷酸酶活性。在完全弗氏佐剂（CFA）炎症性疼痛模型中，单次椎管内注射mPAP蛋白具有持久（三天）的抗痛觉特性，包括抗痛觉过敏和抗异动作用。这些抗伤感受作用被A1R拮抗剂8-环戊基- 1,3 -二丙基黄嘌呤（CPX）短暂阻断，表明mPAP像人和牛PAP一样将核苷酸去磷酸化为腺苷来介导抗伤感受。我们的研究表明，PAP具有物种保守的抗感知作用，并具有ph依赖性的外核苷酸酶活性。以ph依赖性方式代谢核苷酸的能力可能与炎症等组织酸中毒和核苷酸释放发生的情况有关。最后，我们的研究表明重组PAP蛋白可以用于治疗动物模型中的慢性疼痛。
Prostatic acid phosphatase (PAP) is expressed in nociceptive neurons and functions as an ectonucleotidase. When injected intraspinally, the secretory isoforms of human and bovine PAP protein have potent and long-lasting antinociceptive effects that are dependent on A1-adenosine receptor (A1R) activation. In this study, we purified the secretory isoform of mouse (m)PAP using the baculovirus expression system to determine if recombinant mPAP also had antinociceptive properties. We found that mPAP dephosphorylated AMP, and to a much lesser extent, ADP at neutral pH (pH 7.0). In contrast, mPAP dephosphorylated all purine nucleotides (AMP, ADP, ATP) at an acidic pH (pH 5.6). The transmembrane isoform of mPAP had similar pH-dependent ectonucleotidase activity. A single intraspinal injection of mPAP protein had long-lasting (three day) antinociceptive properties, including antihyperalgesic and antiallodynic effects in the Complete Freund's Adjuvant (CFA) inflammatory pain model. These antinociceptive effects were transiently blocked by the A1R antagonist 8-cyclopentyl-1, 3-dipropylxanthine (CPX), suggesting mPAP dephosphorylates nucleotides to adenosine to mediate antinociception just like human and bovine PAP. Our studies indicate that PAP has species-conserved antinociceptive effects and has pH-dependent ectonucleotidase activity. The ability to metabolize nucleotides in a pH-dependent manner could be relevant to conditions like inflammation where tissue acidosis and nucleotide release occur. Lastly, our studies demonstrate that recombinant PAP protein can be used to treat chronic pain in animal models.
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