Cloning of a novel protein interacting with BRS-3 and its effects in wound repair of bronchial epithelial cells.
Cloning of a novel protein interacting with BRS-3 and its effects in wound repair of bronchial epithelial cells.
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与BRS-3相互作用的新型蛋白的克隆及其对支气管上皮细胞损伤修复的影响
DOI：
10.1371/journal.pone.0023072
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发表时间：
2011
期刊：
影响因子：
3.7
通讯作者：
Qin XQ
中科院分区：
文献类型：
--
作者：
Liu HJ;Tan YR;Li ML;Liu C;Xiang Y;Qin XQ
蛙皮素受体亚型3（BRS-3）是一种孤儿蛙皮素受体，可能在肺和气道上皮细胞的应激反应调节中发挥作用。蛙皮素受体激活蛋白（BRAP）是我们在前期研究中发现的一种与BRS-3相互作用的新蛋白。本研究旨在观察BRAP在人支气管上皮细胞（HBECs）中的亚细胞定位及其损伤修复功能。RT-PCR扩增BRAP ORF，连接到pEGFP-C1载体上，将重组质粒pEGFP-C1-BRAP转染Hela细胞。激光共聚焦显微镜观察BRAP蛋白的定位，Western-blot分析BRAP蛋白的表达。同时构建重组质粒pcDNA3.1（+）-BRAP，转染HBECs，观察其对HBECs细胞周期和创伤修复的影响。结果表明，BRAP定位于细胞膜和细胞质，在转染细胞中表达显著增加。流式细胞仪检测结果显示，重组质粒使S期+G2期细胞数增加了25%。显微视频分析系统显示，通过稳定表达BRAP，损伤的HBECs修复指数提高了20%。本研究证实BRAP定位于细胞膜和细胞质中，提示该蛋白是一种细胞质蛋白，可促进HBECs的细胞周期和创伤修复。
Bombesin receptor subtype 3 (BRS-3), the orphan bombesin receptor, may play a role in the regulation of stress responses in lung and airway epithelia. Bombesin receptor activated protein (BRAP )is a novel protein we found in our previous study which interacts with BRS-3. This study was designed to observe the subcellular location and wound repair function of BRAP in human bronchial epithelial cells (HBECs). BRAP ORF was amplified by RT-PCR and ligated to pEGFP-C1 vector, and then the recombinant plasmid pEGFP-C1-BRAP was transfected into Hela cells. The location of BRAP protein was observed by laser confocal microscope, and the expression of it was analyzed by Western-blot. At the same time,we built the recombinant plasmid pcDNA3.1(+)-BRAP, transfected it into HBECs and observed its impact on cell cycle and wound repair of HBECs. The results showed that BRAP locates in membrane and cytoplasm and increases significantly in transfected cells. Flow cytometry results demonstrated that the recombinant plasmid increases S phase plus G2 phase of cell cycle by 25%. Microscopic video analysis system showed that the repair index of wounded HBECs increases by 20% through stable expression of BRAP. The present study demonstrated that BRAP locates in the membrane and cytoplasm, suggesting that this protein is a cytoplasm protein, which promotes cell cycle and wound repair of HBECs.
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DOI：
10.1165/rcmb.2006-0384oc
发表时间：
2007-07-01
影响因子：
6.4
作者：
Gomperts, Brigitte N.;Belperio, John A.;Strieter, Robert M.
通讯作者：
Strieter, Robert M.
DOI：
10.1152/ajplung.00436.2002
发表时间：
2004-01-01
影响因子：
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DOI：
10.1152/ajplung.1995.269.1.l119
发表时间：
1995-07-01
影响因子：
4.9
作者：
KIM, JS;RABE, KF;WHITE, SR
通讯作者：
WHITE, SR
影响因子：
7
作者：
Dunkley, TPJ;Watson, R;Lilley, KS
通讯作者：
Lilley, KS
DOI：
10.1016/j.bbrc.2005.12.084
发表时间：
2006-02-17
影响因子：
3.1
作者：
Wada, K;Tanji, K;Kamitani, T
通讯作者：
Kamitani, T