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首页> 外文期刊>International journal of molecular medicine >'Decoy peptide' region (RIFLKRMPSI) of prorenin prosegment plays a crucial role in prorenin binding to the (pro)renin receptor.
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'Decoy peptide' region (RIFLKRMPSI) of prorenin prosegment plays a crucial role in prorenin binding to the (pro)renin receptor.

机译：肾素原前体的“诱饵肽”区域（RIFLKRMPSI）在肾素原与（肾素原）受体结合中起关键作用。

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摘要

This study investigated a role of decoy peptide region (R10PIFLKRMPSI19P) in prorenin prosegment for prorenin binding to the (pro)renin receptor using the surface plasmon resonance technique. Three kinds of anti-receptor antibodies labeled as anti-107/121, anti-221/235 and anti-His tag antibody were prepared. The respective antigens D107SVANSIHSLFSEET121 (close to the N-terminal side of receptor), E221IGKRYGEDSEQFRD235 (N-terminal side of the transmembrane part of receptor) and 10xHis sequence (C-terminus) were designed based on the sequence of the receptor. These antibodies were immobilized on the CM5 sensor chip by amine coupling and allowed to bind to the receptor. Human prorenin, renin and the decoy bound to the receptor associated with antibodies. Their association (ka) and dissociation (kd) rate constants were measured and the dissociation constants (KD) were determined using Langmuir 1:1 kinetic binding model. The KD for interaction of prorenin and receptor associated to anti-107/121, anti-221/235 and anti-His tag antibodies were 2.9, 1.2 and 7.8 nM, respectively and for renin they were 9.3, 4.4 and 7.1 nM. The decoy bound to the respective immobilized receptor-antibody complexes at KD's of 6.2, 3.5 and 15.2 nM. Prorenin, renin and decoy had lower KD at the nanomolar ranges compared to those of L1PPTD4P in the prorenin prosegment and A248KKRLFDYVV257 in the C-domain of mature renin. The decoy reduced the binding of not only prorenin but also renin to (P)RR. These data are direct evidence that prorenin, renin and the peptides bind to (P)RR and the decoy reduces prorenin binding, supporting our hypothesis that decoy peptide region has a crucial role in prorenin binding.
机译：这项研究使用表面等离子体共振技术研究了诱饵肽区（R10PIFLKRMPSI19P）在prorenin前区中prorenin与（pro）renin受体结合的作用。制备了标记为抗107/121，抗-221/235和抗His标签抗体的三种抗受体抗体。基于受体的序列设计了相应的抗原D107SVANSIHSLFSEET121（靠近受体的N末端侧），E221IGKRYGEDSEQFRD235（受体的跨膜部分的N末端侧）和10xHis序列（C末端）。通过胺偶联将这些抗体固定在CM5传感器芯片上，并使其与受体结合。人肾素原，肾素和诱饵与与抗体相关的受体结合。测量它们的缔合（ka）和解离（kd）速率常数，并使用Langmuir 1：1动力学结合模型确定解离常数（KD）。肾素与抗107/121，抗-221/235和抗His标签抗体相关的受体相互作用的KD分别为2.9、1.2和7.8 nM，而肾素为9.3、4.4和7.1 nM。诱饵以6.2、3.5和15.2nM的KD结合到各自固定的受体-抗体复合物。与在肾素前体部分中的L1PPTD4P和在成熟肾素的C域中的A248KKRLFDYVV257相比，肾上腺素，肾素和诱饵的纳摩尔浓度下的KD较低。诱饵降低了不仅肾素和肾素与（P）RR的结合。这些数据是直接证据，表明肾上腺素，肾素和肽与（P）RR结合，诱饵减少了肾上腺素的结合，支持了我们的假说：诱饵肽区域在肾素中起着至关重要的作用。

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