Live-cell super-resolution imaging of actin using LifeAct-14 with a PAINT-based approach

被引：6

作者：

Bhaskar, Haresh ^{[1
,2
]}

Kleinjan, Dirk-Jan ^{[3
,4
]}

Oi, Curran ^{[5
]}

Gidden, Zoe ^{[2
]}

Rosser, Susan J. J. ^{[3
,4
]}

Horrocks, Mathew H. H. ^{[2
]}

Regan, Lynne ^{[1
]}

机构：

[1] Univ Edinburgh, Sch Biol Sci, Edinburgh, Scotland

[2] Univ Edinburgh, EaStCHEM Sch Chem, Edinburgh, Scotland

[3] Univ Edinburgh, Ctr Synthet & Syst Biol, Sch Biol Sci, Edinburgh, Scotland

[4] Univ Edinburgh, UK Ctr Mammalian Synthet Biol, Sch Biol Sci, Edinburgh, Scotland

[5] Univ Washington, Dept Genome Sci, Seattle, WA USA

来源：

PROTEIN SCIENCE | 2023年 / 32卷 / 02期

基金：

英国生物技术与生命科学研究理事会; 英国惠康基金;

关键词：

actin; LifeAct; live-cell imaging; single-molecule localization microscopy (SMLM); super-resolution (SR) imaging; COMBINATORIAL LIBRARIES; MICROSCOPY; PROTEINS; BINDING; LIMIT;

D O I：

10.1002/pro.4558

中图分类号：

Q5 [生物化学]; Q7 [分子生物学];

学科分类号：

071010 ; 081704 ;

摘要：

We present direct-LIVE-PAINT, an easy-to-implement approach for the nanoscopic imaging of protein structures in live cells using labeled binding peptides. We demonstrate the feasibility of direct-LIVE-PAINT with an actin-binding peptide fused to EGFP, the location of which can be accurately determined as it transiently binds to actin filaments. We show that direct-LIVE-PAINT can be used to image actin structures below the diffraction-limit of light and have used it to observe the dynamic nature of actin in live cells. We envisage a similar approach could be applied to imaging other proteins within live mammalian cells.

引用

页数：8

共 33 条

[1] Structure of the Lifeact-F-actin complex [J].