Trypsin IV or mesotrypsin and p23 cleave protease-activated receptors 1 and 2 to induce inflammation and hyperalgesia
Trypsin IV or mesotrypsin and p23 cleave protease-activated receptors 1 and 2 to induce inflammation and hyperalgesia
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DOI：
10.1074/jbc.m703840200
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发表时间：
2007-09-07
影响因子：
4.8
通讯作者：
Bunnett, Nigel W.
中科院分区：
文献类型：
--
作者：
Knecht, Wolfgang;Cottrell, Graeme S.;Bunnett, Nigel W.
胰蛋白酶主要由胰腺产生，用于降解肠道中的膳食蛋白质，但它也在神经系统和上皮组织中表达，在这些组织中，胰蛋白酶具有多种可由蛋白酶激活受体（PARs）介导的作用。我们检测了人胰蛋白酶IV（或中胰蛋白酶）和大鼠p23的生物学作用，它们是胰蛋白酶的抑制剂抗性形式。酶原胰蛋白酶原IV和pro-p23在大肠杆菌中表达，纯化后具有明显的同质性。肠肽酶裂解两种酶原，释放出对大豆胰蛋白酶抑制剂和抑酶蛋白具有抗性的活性胰蛋白酶IV和p23。胰蛋白酶IV在暴露拴链配体的位点切割PAR(1)、PAR(2)和PAR(4)的n端片段(PAR(1) = PAR（4) > PAR(2)）。胰蛋白酶IV在表达人PAR(1)和PAR(2)的转染细胞中增加[Ca2(+)] (i)，具有相似的效力(PAR(1), 0.5 μ M；PAR(2), 0.6 μ M). p23也裂解PAR(1)和PAR(2)片段，并向表达这些受体的细胞发出信号。胰蛋白酶IV和p23增加了对辣椒素有反应的大鼠背根神经节神经元中的[Ca2(+)] i，从而介导神经源性炎症和伤害感觉。小鼠足底注射胰蛋白酶IV和p23可诱导小鼠水肿和粒细胞浸润，而PAR(1)(-/-)（胰蛋白酶IV）和PAR(2)(-/-)（胰蛋白酶IV和p23）小鼠无此现象。胰蛋白酶IV和p23在小鼠中引起热痛觉过敏和机械异常性痛觉过敏，这些作用在PAR(2)（-/-）小鼠中不存在，但在PAR1（-/-）小鼠中维持。因此，胰蛋白酶IV和p23是抑制剂抗性胰蛋白酶，它们可以切割和激活PAR，引起PAR(1)-和PAR(2)-依赖性炎症和PAR(2)-依赖性痛觉过敏。
Although principally produced by the pancreas to degrade dietary proteins in the intestine, trypsins are also expressed in the nervous system and in epithelial tissues, where they have diverse actions that could be mediated by protease-activated receptors ( PARs). We examined the biological actions of human trypsin IV ( or mesotrypsin) and rat p23, inhibitor-resistant forms of trypsin. The zymogens trypsinogen IV and pro-p23 were expressed in Escherichia coli and purified to apparent homogeneity. Enteropeptidase cleaved both zymogens, liberating active trypsin IV and p23, which were resistant to soybean trypsin inhibitor and aprotinin. Trypsin IV cleaved N-terminal fragments of PAR(1), PAR(2), and PAR(4) at sites that would expose the tethered ligand (PAR(1) = PAR(4) > PAR(2)). Trypsin IV increased [Ca2(+)] (i) in transfected cells expressing human PAR(1) and PAR(2) with similar potencies (PAR(1), 0.5 mu M; PAR(2), 0.6 mu M). p23 also cleaved fragments of PAR(1) and PAR(2) and signaled to cells expressing these receptors. Trypsin IV and p23 increased [Ca2(+)] i in rat dorsal root ganglion neurons that responded to capsaicin and which thus mediate neurogenic inflammation and nociception. Intraplantar injection of trypsin IV and p23 in mice induced edema and granulocyte infiltration, which were not observed in PAR(1) (-/-)(trypsin IV) and PAR(2) (-/-) ( trypsin IV and p23) mice. Trypsin IV and p23 caused thermal hyperalgesia and mechanical allodynia and hyperalgesia in mice, and these effects were absent in PAR(2) (-/-) mice but maintained in PAR1 (-/-) mice. Thus, trypsin IV and p23 are inhibitor- resistant trypsins that can cleave and activate PARs, causing PAR(1)- and PAR(2)- dependent inflammation and PAR(2)- dependent hyperalgesia.