Novel ecto-tagged integrins reveal their trafficking in live cells

被引：40

作者：

Huet-Calderwood, Clotilde ^{[1
]}

Rivera-Molina, Felix ^{[2
]}

Iwamoto, Daniel V. ^{[1
]}

Kromann, Emil B. ^{[2
,3
]}

Toomre, Derek ^{[2
]}

Calderwood, David A. ^{[1
,2
]}

机构：

[1] Yale Univ, Sch Med, Dept Pharmacol, 333 Cedar St, New Haven, CT 06520 USA

[2] Yale Univ, Sch Med, Dept Cell Biol, 333 Cedar St, New Haven, CT 06520 USA

[3] Yale Univ, Dept Biomed Engn, 333 Cedar St, New Haven, CT 06520 USA

来源：

NATURE COMMUNICATIONS | 2017年 / 8卷

基金：

英国惠康基金; 美国国家卫生研究院;

关键词：

CRYSTAL-STRUCTURE; CYTOPLASMIC TAIL; EXTRACELLULAR SEGMENT; STRUCTURAL BASIS; ACTIVATION; ALPHA-V-BETA-3; ECTODOMAIN; ADHESION; COMPLEX; LIGAND;

D O I：

10.1038/s41467-017-00646-w

中图分类号：

O [数理科学和化学]; P [天文学、地球科学]; Q [生物科学]; N [自然科学总论];

学科分类号：

07 ; 0710 ; 09 ;

摘要：

Integrins are abundant heterodimeric cell-surface adhesion receptors essential in multicellular organisms. Integrin function is dynamically modulated by endo-exocytic trafficking, however, major mysteries remain about where, when, and how this occurs in living cells. To address this, here we report the generation of functional recombinant beta 1 integrins with traceable tags inserted in an extracellular loop. We demonstrate that these 'ecto-tagged' integrins are cell-surface expressed, localize to adhesions, exhibit normal integrin activation, and restore adhesion in beta 1 integrin knockout fibroblasts. Importantly, beta 1 integrins containing an extracellular pH-sensitive pHluorin tag allow direct visualization of integrin exocytosis in live cells and revealed targeted delivery of integrin vesicles to focal adhesions. Further, using beta 1 integrins containing a HaloTag in combination with membrane-permeant and -impermeant Halo dyes allows imaging of integrin endocytosis and recycling. Thus, ecto-tagged integrins provide novel powerful tools to characterize integrin function and trafficking.

引用

页数：13

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