Localization of human (pro)renin receptor lacking the transmembrane domain on budded baculovirus of Autographa californica multiple nucleopolyhedrovirus
Localization of human (pro)renin receptor lacking the transmembrane domain on budded baculovirus of Autographa californica multiple nucleopolyhedrovirus
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DOI：
10.1007/s00253-008-1776-8
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发表时间：
2009-03-01
影响因子：
5
通讯作者：
Park, Enoch Y.
中科院分区：
文献类型：
--
作者：
Kato, Tatsuya;Du, Dongning;Park, Enoch Y.
利用昆虫细胞表达了具有天然跨膜和胞浆结构域的人(PRO)肾素受体(hPRR-WTM)和缺乏两者的hPRR(hPRR-w/OTM)。在感染的SF-9细胞中，hPRR-WTM表达于细胞核的周边区域，并定位于内质网(ER)。然而，在4A℃下用Triton X-100处理不能从重组链球绦虫多核型多角体病毒(AcMNPV)中提取出hPRR-w/OTM，相反，hPRR-w/OTM存在于感染的SF-9细胞胞浆的点状结构域中，但细胞内hPRR-w/OTM不共定位于高尔基体和溶酶体内。这表明hPRR-WTM和hPRR-w/OTM分别定位于SF-9细胞的内质网和细胞质。此外，Western blotting证实hPRR-w/OTM在重组AcMNPV芽生杆状病毒中的定位。这是第一次发现缺乏跨膜结构域的外来蛋白与杆状病毒的关联。如果这一发现适用于双展示系统，能够在杆状病毒包膜和衣壳上表达，将为组织和细胞特异性靶向和细胞内靶向的杆状病毒展示系统提供新的方法学。
Human (pro)renin receptor (hPRR), a construct with native transmembrane and cytoplasmic domains (hPRR-wTM), and hPRR lacking both (hPRR-w/oTM) were expressed using insect cells. The hPRR-wTM was expressed in the peripheral domains of the nucleus in infected Sf-9 cells, and its localization was observed in endoplasmic reticulum (ER). However, it could not be extracted from recombinant Autographa californica multiple nucleopolyhedrovirus (AcMNPV) by Triton X-100 treatment at 4A degrees C. In contrast, hPRR-w/oTM was observed in punctate domains in the cytoplasm of infected Sf-9 cells, but intracellular hPRR-w/oTM did not co-localize in the Golgi apparatus and lysosomes. This indicates that hPRR-wTM and hPRR-w/oTM is localized in the ER and cytoplasmic organelles of Sf-9 cell, respectively. Moreover, the localization of hPRR-w/oTM in budded baculovirus of recombinant AcMNPV was confirmed by Western blotting. This is the first finding of the association of a foreign protein lacking a transmembrane domain with a baculovirus. If this finding is available for double displaying system, being capable of expression on the envelope and the capsid of baculovirus, it will lead to new methodology of baculovirus display system for tissue- and cell-specific targeting and intracellular targeting.