Four day inhibition of prolyl oligopeptidase causes significant changes in the peptidome of rat brain, liver and kidney
Four day inhibition of prolyl oligopeptidase causes significant changes in the peptidome of rat brain, liver and kidney
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DOI：
10.1016/j.biochi.2012.04.005
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发表时间：
2012-09-01
期刊：
影响因子：
3.9
通讯作者：
Garcia-Horsman, J. Arturo
中科院分区：
文献类型：
--
作者：
Tenorio-Laranga, Jofre;Mannisto, Pekka T.;Garcia-Horsman, J. Arturo
脯氨酰寡肽酶 (PREP) 可在脯氨酸的 C 侧切割短肽。尽管几种含有脯氨酸的神经肽已被证明可以在体外被 PREP 有效切割，但这种肽酶的实际生理底物仍然存在争议。本研究的目的是使用我们最近开发的基于 iTRAQ 的技术，评估强效特异性 PREP 抑制剂 KYP-2047 重复 4 天给药所引起的大鼠组织肽组的变化。我们发现主要源自胞质蛋白的特定肽亚群的水平发生组织依赖性变化。特别是在肾脏中，细胞色素c氧化酶的水平下降，许多改变的肽源自参与能量代谢的线粒体蛋白。然而，在下丘脑中，我们发现前激素前体肽衍生的显着变化。我们无法确认 PREP 作为 β-内啡肽、甘丙肽、十八烷神经肽、神经肽-谷氨酸-异亮氨酸、P 物质、生长抑素、脑啡肽和神经肽 Y 的代谢酶的作用。此外，其中一些神经肽以及大多数源自其他较大蛋白质的神经肽的降解模式的变化并不遵循脯氨酸的特异性。经过 4 天的处理后，我们发现大量肽（全部源自分泌的前蛋白）被一对基本残基特异性裂解。体外实验表明，在 KYP-2047 敏感的 v/ay 中，PREP 修饰内源二碱基残基特异性蛋白水解。这些发现表明，PREP 可能在导致我们观察到的特定肽变化的途径中间接发挥作用。这里报告的数据表明 l'REP 具有更广泛的组织特异性生理作用，而不仅仅是含有活性肽和激素的脯氨酸的代谢。 (C) 2012 Elsevier Masson SAS。版权所有。
Prolyl oligopeptidase (PREP) cleaves short peptides at the C-side of proline. Although several proline containing neuropeptides have been shown to be efficiently cleaved by PREP in vitro, the actual physiological substrates of this peptidase are still a matter of controversy. The aim of this study was to evaluate the changes in the peptidome of rat tissues caused by a repeated 4-day administration of the potent and specific PREP inhibitor KYP-2047, using our recently developed iTRAQ-based technique. We found tissue-dependent changes in the levels of specific subsets of peptides mainly derived from cytosolic proteins. Particularly in the kidney, where the levels of cytochrome c oxidase were found decreased, many of the altered peptides originated from mitochondrial proteins being involved in energy metabolism. However, in the hypothalamus, we found significant changes in peptides derived front hormone precursors. We could not confirm a role of PREP as the metabolising enzyme for beta-endorphin, galanin, octadecaneuropeptide, neuropeptide-glutamic acid-isoleucine, substance P. somatostatin, enkephalin and neuropeptide Y. Furthermore, changes in the degradation patterns of some of these neuropeptides, and also most of those derived from other larger proteins, did not follow specificity to proline. After a 4-day treatment, we found a significant amount of peptides, all derived from secreted pro-proteins, being cleaved with pair of basic residue specificity. In vitro experiments indicated that PREP modifies the endogenous dibasic residue specific proteolysis, in a KYP-2047 sensitive v/ay. These findings suggest that PREP may act indirectly within the routes leading to the specific peptide changes that we observed. The data reported here suggest a wider tissue specific physiological role of l'REP rather than the mere metabolism of proline containing active peptides and hormones. (C) 2012 Elsevier Masson SAS. All rights reserved.