Search of the human proteome for endomorphin-1 and endomorphin-2 precursor proteins.
Search of the human proteome for endomorphin-1 and endomorphin-2 precursor proteins.
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在人类蛋白质组中搜索内吗啡肽 1 和内吗啡肽 2 前体蛋白。
DOI：
10.1016/j.lfs.2007.09.025
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发表时间：
2007
期刊：
影响因子：
6.1
通讯作者：
Howells,RichardD
中科院分区：
文献类型：
--
作者：
Terskiy,Alexandra;Wannemacher,KennethM;Yadav,PremN;Tsai,Michael;Tian,Bin;Howells,RichardD
基于肽Tyr-Pro-Trp-Gly-NH 2（Tyr-W-MIF）的有前景的阿片样物质药理学特征，Zadina等人[Zadina，J.E.，哈克勒湖，盖湖J.，Kastin，A.J.，1997.μ-阿片受体的有效和选择性内源性激动剂。Nature 386，499-5502]合成并筛选了其他Gly 4取代的肽，最终合成了Tyr-Pro-Trp-Phe-NH 2（内吗啡肽-1），其对μ-阿片受体显示出高亲和力和选择性。然后从牛脑额叶皮层中分离出酰胺化肽，以及一种相关肽Tyr-Pro-Phe-Phe-NH 2（内吗啡肽-2），其对μ-阿片受体表现出类似的高亲和力和选择性。内吗啡肽在脑中的生物合成仍然不清楚，因为肽的推定前体蛋白尚未被鉴定。随着人类基因组测序计划的完成，我们假设我们应该使用生物信息学方法来发现肽的生物前体，以在当前的人类蛋白质组中搜索包含内吗啡肽序列以及随后的Gly-Lys/Arg（肽α-酰胺化和前体切割的共有序列）的蛋白质。12个蛋白质被鉴定含有内吗啡肽-1 Tyr-Pro-Trp-Phe序列，但是没有一个蛋白质含有α-酰胺化所必需的Tyr-Pro-Trp-Phe-Gly序列。22种不同的蛋白质含有内吗啡肽-2四肽序列，其中两种含有Tyr-Pro-Phe-Phe-Gly序列，然而，没有一种含有必需的肽-Gly-Lys/Arg序列。使用内吗啡肽-2抗体的蛋白质印迹分析检测到4个突出的蛋白质在小鼠大脑中，需要重新解释以前的免疫细胞定位研究在大脑中。目前人类蛋白质组的筛选没有得到内吗啡肽前体蛋白的证据，根据公认的生化标准。
Based on the promising opioid pharmacological profile of the peptide, Tyr-Pro-Trp-Gly-NH2(Tyr-W-MIF), Zadina et al. [Zadina, J.E., Hackler, L., Ge, L.-J., Kastin, A.J., 1997. A potent and selective endogenous agonist for the μ-opiate receptor. Nature 386, 499–5502] synthesized and screened other Gly4-substituted peptides, culminating in the synthesis of Tyr-Pro-Trp-Phe-NH2(endomorphin-1), which displayed high affinity and selectivity for the μ-opioid receptor. The amidated peptide was then isolated from bovine brain frontal cortex, as was a related peptide, Tyr-Pro-Phe-Phe-NH2(endomorphin-2), that displayed similar high affinity and selectivity for the μ-opioid receptor. The biosynthesis of the endomorphins in the brain remains obscure, since the putative precursor proteins for the peptides have not been identified. With the completion of the human genome sequencing project, we hypothesized that we should uncover the biological precursors of the peptides using a bioinformatic approach to search the current human proteome for proteins that contained the endomorphin peptide sequences followed by Gly-Lys/Arg, the consensus sequence for peptide α-amidation and precursor cleavage. Twelve proteins were identified that contained the endomorphin-1 Tyr-Pro-Trp-Phe sequence, however none contained the Tyr-Pro-Trp-Phe-Gly sequence necessary for α-amidation. Twenty-two distinct proteins contained the endomorphin-2 tetrapeptide sequence, and two of those contained the sequence, Tyr-Pro-Phe-Phe-Gly, however, none contained the requisite peptide-Gly-Lys/Arg sequence. Western blot analysis using an endomorphin-2 antibody detected 4 prominent proteins in mouse brain, necessitating reinterpretation of previous immunocytolocalization studies in the brain. Screening of the current human proteome yielded no evidence for endomorphin precursor proteins based on accepted biochemical criteria.