Muted蛋白介导CD63在嗜铬细胞大致密核粒的定位

doi:10.16288/j.yczz.16-156

遗传 ›› 2016, Vol. 38 ›› Issue (8): 718-723.doi: 10.16288/j.yczz.16-156

Muted蛋白介导CD63在嗜铬细胞大致密核粒的定位

郝振华, 李巍

首都医科大学附属北京儿童医院,北京市儿科研究所医学遗传中心,教育部儿科重大疾病研究重点实验室,北京 100045

收稿日期:2016-05-03 修回日期:2016-06-06 出版日期:2016-08-20 发布日期:2016-06-21
通讯作者: 李巍,博士,教授,研究方向：医学遗传学与细胞生物学。E-mail: liwei@bch.com.cn E-mail:nanshayu@163.com
作者简介:郝振华,博士,助理研究员,研究方向：医学遗传学与细胞生物学。E-mail: nanshayu@163.com
基金资助:
北京市自然科学基金项目(编号：5164032)资助

Muted protein is involved in the targeting of CD63 to large dense-core vesicles of chromaffin cells

Zhenhua Hao, Wei Li

Key Laboratory of Major Diseases in Children, Ministry of Education, Center for Medical Genetics, Beijing Pediatric Research Institute, Beijing Children’s Hospital, Capital Medical University, Beijing 100045, China

Received:2016-05-03 Revised:2016-06-06 Online:2016-08-20 Published:2016-06-21
Supported by:
Supported by Beijing Natural Science Foundation (No; 5164032)

摘要/Abstract

摘要： 大致密核心颗粒(Large dense-core vesicles,LDCVs)是一种溶酶体相关细胞器(Lysosome-related organelles,LROs),在细胞受到刺激时快速释放其内含物,从而调节机体生长发育、物质代谢和能量代谢等,维持机体的稳态。Muted蛋白是溶酶体相关细胞器生物发生复合体-1(Biogenesis of lysosomal organelles complex-1,BLOC-1)的一个亚基,参与调控溶酶体和多种细胞特异性LROs的生物学发生。四联体跨膜蛋白CD63最初被定位在内体-溶酶体系统,后来发现它也参与部分LROs膜的组成。CD63是否存在于LDCVs尚不清楚,其靶向运输过程是否依赖Muted蛋白也不明确。本研究以肾上腺嗜铬细胞为细胞模型,采用荧光共定位、活细胞追踪和密度梯度离心等实验鉴定CD63蛋白为LDCVs的膜组分,并探讨了其生物学功能。活细胞实验显示CD63-YFP特异性定位在NPY-dsRed标记的LDCVs上,并动态参与LDCVs膜的组成;密度梯度离心实验表明高密度区的CD63与LDCVs的标记蛋白VMAT1共同出峰;Muted蛋白缺乏的小鼠(Bloc1s5基因突变)是一种理想的Hermansky-Pudlak综合征(HPS)小鼠模型, 免疫印迹实验显示该突变体小鼠肾上腺组织中CD63蛋白含量明显减少,暗示Muted蛋白可能参与CD63的分选。以上结果表明CD63是LDCVs的膜成分,CD63在胞内的稳态水平依赖于Muted蛋白,为HPS的病理发生机制提供一定的理论依据。

关键词: 大致密核心颗粒, CD63, Muted, 溶酶体相关细胞器, Hermansky-Pudlak综合征

Abstract: Large dense-core vesicles (LDCVs) are characterized as a class of lysosome-related organelles (LROs), which undergo regulated release and play important roles in development, metabolism and homeostasis. The Muted protein is a subunit of the biogenesis of lysosome-related organelles complex-1 (BLOC-1), which functions in the biogenesis of lysosomes and LROs. CD63 is a membrane component of lysosomes and LROs. Whether and how CD63 is sorted into LDCVs is largely unknown. In this study, we aim to identify the localization of CD63 in chromaffin cells by colocalization, living cell imaging and cell fractionation. We found that a proportion of CD63-YFP colocalized with NPY-dsRed labeled LDCVs. By sucrose density gradient fractionation, a proportion of CD63 was found to be highly enriched in LDCVs fractions. The Muted mutant mouse is a model of Hermansky-Pudlak syndrome (HPS). We also found that the level of CD63 was significantly decreased in Muted-deficient adrenal glands, suggesting that the Muted protein is important for the steady-state level of CD63. Our results suggest that CD63 is a membrane component of LDCVs and the stability of CD63 is dependent on the Muted protein, which provides a clue to the pathogenesis of LRO defects in HPS.

Key words: large dense-core vesicles, CD63, Muted, lysosome-related organelle, Hermansky-Pudlak syndrome

郝振华, 李巍. Muted蛋白介导CD63在嗜铬细胞大致密核粒的定位[J]. 遗传, 2016, 38(8): 718-723.

Zhenhua Hao, Wei Li. Muted protein is involved in the targeting of CD63 to large dense-core vesicles of chromaffin cells[J]. Hereditas(Beijing), 2016, 38(8): 718-723.

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Muted蛋白介导CD63在嗜铬细胞大致密核粒的定位

Muted protein is involved in the targeting of CD63 to large dense-core vesicles of chromaffin cells

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[1]	李泰明, 蓝文俊, 黄灿, 张春, 刘晓玫. 近红外荧光蛋白标记乳腺癌细胞外泌体的构建及鉴定[J]. 遗传, 2016, 38(5): 427-435.
[2]	张喆，李巍. Weibel-Palade小体形成和功能研究进展[J]. 遗传, 2009, 31(9): 882-888.