Advances in analysis of low signal-to-noise images link dynamin and AP2 to the functions of an endocytic checkpoint.
Advances in analysis of low signal-to-noise images link dynamin and AP2 to the functions of an endocytic checkpoint.
复制标题
DOI：
10.1016/j.devcel.2013.06.019
复制
发表时间：
2013-08-12
影响因子：
11.8
通讯作者：
Danuser, Gaudenz
中科院分区：
文献类型：
--
作者：
Aguet, Francois;Antonescu, Costin N.;Mettlen, Marcel;Schmid, Sandra L.;Danuser, Gaudenz
许多内吞辅助蛋白（EAP）介导网格蛋白包被的小凹（CCP）组装和成熟为含货物的囊泡。由于EAP之间的潜在冗余，我们在这里展示的网格蛋白介导的内吞作用（CME）的可塑性和弹性，通过批量测量货物摄取EAP功能的分析已受到阻碍。相反，EAP功能的最佳研究是通过揭示EAP与个体CCP之间的相关性以及由此产生的成熟变化。然而，大多数EAP与CCP结合的数量很少，使得通过融合的荧光报告物测量EAP缔合非常容易受到检测错误的影响。在这里，我们提出了一个框架，无偏测量EAP招聘CCP和CCP动态的直接影响。我们鉴定了发动蛋白和AP 2接头的EAP结合α-adaptin附属结构域作为受调控的多步成熟过程中的开关，并为CME中的分子检查点提供了直接证据。
Numerous endocytic accessory proteins (EAPs) mediate assembly and maturation of clathrin-coated pits (CCPs) into cargo-containing vesicles. Analysis of EAP function through bulk measurement of cargo uptake has been hampered due to potential redundancy among EAPs and, as we show here, the plasticity and resilience of clathrin-mediated endocytosis (CME). Instead, EAP function is best studied by uncovering the correlation between variations in EAP association to individual CCPs and the resulting variations in maturation. However, most EAPs bind to CCPs in low numbers, making the measurement of EAP association via fused fluorescent reporters highly susceptible to detection errors. Here, we present a framework for unbiased measurement of EAP recruitment to CCPs and their direct effects on CCP dynamics. We identify dynamin and the EAP-binding α-adaptin appendage domain of the AP2 adaptor as switches in a regulated, multistep maturation process and provide direct evidence for a molecular checkpoint in CME.
登录
查看更多内容
影响因子：
9.8
作者：
Loerke D;Mettlen M;Yarar D;Jaqaman K;Jaqaman H;Danuser G;Schmid SL
通讯作者：
Schmid SL
影响因子：
3.3
作者：
Antonescu CN;Aguet F;Danuser G;Schmid SL
通讯作者：
Schmid SL
影响因子：
64.5
作者：
Puthenveedu, Manojkumar A.;von Zastrow, Mark
通讯作者：
von Zastrow, Mark
涂有网状蛋白涂层坑的前五秒钟。
影响因子：
64.5
作者：
Cocucci E;Aguet F;Boulant S;Kirchhausen T
通讯作者：
Kirchhausen T
影响因子：
21.3
作者：
Merrifield, CJ;Feldman, ME;Almers, W
通讯作者：
Almers, W