利用 Bm 细胞在 Bombyx mori 核型多角体病毒（BmNPV）颗粒上展示人（pro）肾素受体。

Display of the human (pro)renin receptor on Bombyx mori nucleopolyhedrovirus (BmNPV) particles using Bm cells.

机构信息

Laboratory of Biotechnology, Faculty of Agriculture, Shizuoka University, 836 Ohya, Suruga-ku, Shizuoka 422-8529, Japan.

出版信息

J Biosci Bioeng. 2012 Nov;114(5):564-9. doi: 10.1016/j.jbiosc.2012.06.008. Epub 2012 Jul 3.

DOI:10.1016/j.jbiosc.2012.06.008

PMID:22762973

Abstract

The human (pro)renin receptor (hPRR) was displayed on the surface of Bombyx mori nucleopolyhedrovirus (BmNPV) with and without fusion to glycoprotein 64 (GP64) of the BmNPV. hPRR1 is a native hPRR with an additional FLAG peptide sequence inserted between the signal peptide and prorenin-binding domain. hPRR2 has the prorenin-binding domain inserted between amino acid residues (81)Asp and (82)Pro of GP64. hPRR4 has the prorenin-binding domain inserted in (81)Asp and (320)Met of partially deleted GP64. Incorporation of hPRR was confirmed in recombinant BmNPV (rBmNPV) but not in cysteine protease-deleted rBmNPV. hPRR1 was observed in ER, but hPRR2 and hPRR4 were observed around the endoplasmic reticulum (ER) and in its periphery. rBmNPV-hPRR1 and -hPRR2, carrying hPRR1 and hPRR2 respectively, showed binding affinity to human renin, but rBmNPV-hPRR4 did not. The presence of hPRR4 of rBmNPV-hPRR4 was confirmed in western blotting under nonreducing conditions, suggesting that although hPRR4 was incorporated in rBmNPV-hPRR4, it behaved as a non-functional aggregate. This rBmNPV display system can also be used for analyzing a ligand-receptor interaction.

摘要

人（前）肾素受体（hPRR）与糖蛋白 64（GP64）融合和不融合展示在桑蚕核型多角体病毒（BmNPV）表面。hPRR1 是一种天然的 hPRR，在信号肽和前肾素结合域之间插入了一个 FLAG 肽序列。hPRR2 在 GP64 的氨基酸残基（81）Asp 和（82）Pro 之间插入了前肾素结合域。hPRR4 在部分缺失的 GP64 的（81）Asp 和（320）Met 之间插入了前肾素结合域。重组 BmNPV（rBmNPV）中证实了 hPRR 的掺入，但在半胱氨酸蛋白酶缺失的 rBmNPV 中没有。hPRR1 在 ER 中观察到，但 hPRR2 和 hPRR4 观察到在 ER 周围及其周围。分别携带 hPRR1 和 hPRR2 的 rBmNPV-hPRR1 和 -hPRR2 显示出与人肾素的结合亲和力，但 rBmNPV-hPRR4 没有。在非还原条件下的 Western blot 中证实了 rBmNPV-hPRR4 中 hPRR4 的存在，表明尽管 hPRR4 被掺入 rBmNPV-hPRR4 中，但它表现为无功能的聚集体。这种 rBmNPV 展示系统也可用于分析配体-受体相互作用。