Characterization of the early endosome and putative endocytic carrier vesicles in vivo and with an assay of vesicle fusion in vitro.

内体 内吞作用 小泡 细胞生物学 脂质双层融合 生物 内吞循环 囊泡融合 诺可达唑 微管 生物化学 突触小泡 细胞骨架 细胞 细胞内
作者
Jean Grüenberg,Gareth Griffiths,K E Howell
出处
期刊:Journal of Cell Biology [The Rockefeller University Press]
卷期号:108 (4): 1301-1316 被引量:584
标识
DOI:10.1083/jcb.108.4.1301
摘要

We have investigated two aspects of membrane traffic at early stages of endocytosis: membrane fusion and microtubule-dependent transport. As a marker, we have used the trans-membrane glycoprotein G of vesicular stomatitis virus implanted into the plasma membrane and then internalized for different times at 37 degrees C. The corresponding endosomal fractions were immunoisolated using the cytoplasmic domain of the G protein as antigen. These fractions were then used in an in vitro assay to quantify the efficiency of fusion between endosomal vesicles. To identify the vesicular partners of the fusion, these in vitro studies were combined with in vivo biochemical and morphological experiments. Internalized molecules were delivered to early endosomal elements, which corresponded to a network of tubular and tubulovesicular structures. Rapid recycling back to the plasma membrane and routing to late stages of the pathway occurred from these early endosomal elements. These elements exhibited a high and specific fusion activity with each other in vitro, suggesting that individual elements of the early endosomal compartment interact with each other in vivo. After their appearance in the early endosome, the molecules destined to be degraded were observed at the next stage of the pathway in distinct spherical vesicles (0.5 micron diam) and then in late endosomes and lysosomes. When the microtubules were depolymerized with nocodazole, endocytosis proceeded as in control cells. However, internalized molecules remained in the spherical vesicles and did not appear in late endosomes or lysosomes. These spherical vesicles had relatively little fusion activity with each other or with early endosomal elements in vitro. Our observations suggest that the spherical vesicles mediate transport between the early endosome and late endosomes and that this process requires intact microtubules.
最长约 10秒,即可获得该文献文件

科研通智能强力驱动
Strongly Powered by AbleSci AI
更新
大幅提高文件上传限制,最高150M (2024-4-1)

科研通是完全免费的文献互助平台,具备全网最快的应助速度,最高的求助完成率。 对每一个文献求助,科研通都将尽心尽力,给求助人一个满意的交代。
实时播报
金有财发布了新的文献求助10
1秒前
1秒前
wang5945发布了新的文献求助10
1秒前
无事小神仙完成签到,获得积分10
2秒前
SciGPT应助边伯贤采纳,获得10
2秒前
2秒前
2秒前
3秒前
Dou完成签到,获得积分10
5秒前
5秒前
5秒前
7秒前
7秒前
7秒前
7秒前
程住气完成签到 ,获得积分10
8秒前
榆木完成签到 ,获得积分10
9秒前
10秒前
11秒前
超级发布了新的文献求助10
13秒前
14秒前
不安的夜柳完成签到,获得积分10
15秒前
15秒前
姜建正完成签到,获得积分20
16秒前
金有财完成签到,获得积分10
17秒前
rnf完成签到,获得积分10
18秒前
YF完成签到 ,获得积分10
18秒前
QJQ完成签到 ,获得积分10
20秒前
爆米花应助想毕业的小李采纳,获得10
21秒前
wyx完成签到 ,获得积分10
22秒前
Azyyyy完成签到,获得积分10
24秒前
winew完成签到 ,获得积分10
25秒前
丘比特应助air采纳,获得10
26秒前
Lucas应助科研通管家采纳,获得10
27秒前
李健应助科研通管家采纳,获得10
27秒前
小马甲应助科研通管家采纳,获得10
27秒前
搜集达人应助科研通管家采纳,获得10
27秒前
27秒前
斯文败类应助科研通管家采纳,获得10
27秒前
Jasper应助科研通管家采纳,获得10
27秒前
高分求助中
Sustainability in Tides Chemistry 2800
The Young builders of New china : the visit of the delegation of the WFDY to the Chinese People's Republic 1000
Rechtsphilosophie 1000
Bayesian Models of Cognition:Reverse Engineering the Mind 888
Le dégorgement réflexe des Acridiens 800
Defense against predation 800
Very-high-order BVD Schemes Using β-variable THINC Method 568
热门求助领域 (近24小时)
化学 医学 生物 材料科学 工程类 有机化学 生物化学 物理 内科学 纳米技术 计算机科学 化学工程 复合材料 基因 遗传学 催化作用 物理化学 免疫学 量子力学 细胞生物学
热门帖子
关注 科研通微信公众号,转发送积分 3136281
求助须知:如何正确求助?哪些是违规求助? 2787312
关于积分的说明 7780922
捐赠科研通 2443313
什么是DOI,文献DOI怎么找? 1299106
科研通“疑难数据库(出版商)”最低求助积分说明 625325
版权声明 600905