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首页> 外文期刊>Biochemistry >Ligand binding-dependent limited proteolysis of the atrial natriuretic peptide receptor: juxtamembrane hinge structure essential for transmembrane signal transduction.
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Ligand binding-dependent limited proteolysis of the atrial natriuretic peptide receptor: juxtamembrane hinge structure essential for transmembrane signal transduction.

机译:心房利钠肽受体的配体结合依赖性有限蛋白水解:跨膜信号转导必不可少的近膜铰链结构。

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摘要

The atrial natriuretic peptide (ANP) receptor is a 130-kDa transmembrane protein containing an extracellular ANP-binding domain, a single transmembrane sequence, an intracellular kinase-homologous domain, and a guanylate cyclase (GCase) domain. We observed that the receptor, when bound with ANP, was rapidly cleaved by endogenous or exogenously added protease to yield a 65-kDa ANP-binding fragment. No cleavage occurred without bound ANP. This ligand-induced cleavage abolished GCase activation by ANP. Cleavage occurred in an extracellular, juxtamembrane region containing six closely spaced Pro residues and a disulfide bond. Such structural features are shared among the A-type and B-type ANP receptors but not by ANP clearance receptors. The potential role of the hinge structure was examined by mutagenesis experiments. Mutation of Pro(417), but not other Pro residues, to Ala abolished GCase activation by ANP. Elimination of the disulfide bond by Cys to Ser mutations yielded a constitutively active receptor. Pro(417), and Cys(423) and Cys(432) forming the disulfide bond are strictly conserved among GCase-coupled receptors, while other residues are largely variable. The conserved Pro(417) and the disulfide bond may represent a consensus signaling motif in the juxtamembrane hinge structure that undergoes a marked conformational change upon ligand binding and apparently mediates transmembrane signal transduction.
机译:心钠素(ANP)受体是一个130 kDa跨膜蛋白,包含一个胞外ANP结合域,一个跨膜序列,一个胞内激酶同源域和一个鸟苷酸环化酶(GCase)域。我们观察到,当受体与ANP结合时,会被内源或外源添加的蛋白酶迅速裂解,产生65 kDa ANP结合片段。没有结合的ANP没有发生切割。该配体诱导的切割消除了ANP对GCase的活化。切割发生在细胞外的近膜区域,该区域包含六个紧密间隔的Pro残基和一个二硫键。此类结构特征在A型和B型ANP受体之间共有,但ANP清除受体则不具备。通过诱变实验检查了铰链结构的潜在作用。 Pro(417),但不是其他Pro残基突变为Ala废除了ANP的GCase激活。 Cys至Ser突变消除了二硫键,产生了组成型活性受体。 Pro(417)以及形成二硫键的Cys(423)和Cys(432)在GCase偶联受体之间严格守恒,而其他残基则在很大程度上可变。保守的Pro(417)和二硫键可能代表了近膜铰链结构中的共有信号基序,该基序在配体结合后发生显着的构象变化,并明显介导跨膜信号转导。

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