Visualizing dynamic interaction between calmodulin and calmodulin-related kinases via a monitoring method in live mammalian cells

被引：16

作者：

Lee, Kyoung Hu ^{[1
]}

Lee, Sangkyu ^{[1
]}

Lee, Woo Yong ^{[1
]}

Yang, Hee Won ^{[1
]}

Heo, Won Do ^{[1
,2
,3
]}

机构：

[1] Korea Adv Inst Sci & Technol, Dept Biol Sci, Taejon 305701, South Korea

[2] Korea Adv Inst Sci & Technol, Grad Sch Nanosci & Technol WCU, Taejon 305701, South Korea

[3] Korea Adv Inst Sci & Technol, KAIST Inst BioCentury KIB, Taejon 305701, South Korea

来源：

PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA | 2010年 / 107卷 / 08期

关键词：

calmodulin-dependent kinase; live cell imaging; Protein-Protein interaction; calcium; small GTPase; DEPENDENT PROTEIN-KINASE; SYNAPTIC PLASTICITY; LIVING CELLS; CALCIUM; ACTIVATION; IV; BIOSENSORS; MEMBRANE; DESIGN; FRET;

D O I：

10.1073/pnas.0911262107

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：

A new visualizing method was developed for monitoring proteinprotein (P-P) interactions in live mammalian cells. P-P interactions are visualized by directing localization of a bait protein to endosomes. This method is sufficiently robust to analyze signal-dependent P-P interactions such as calcium-dependent protein interactions. We visualized interactions between activated calmodulin and calmodulin-binding proteins, and observed oscillatory interactions via time-lapse imaging. In addition, this new method can simultaneously monitor multiple P-P interactions in a single live cell, which allows comparison of interactions between several prey proteins and a single bait protein. We observed that CaMKK1 and CaMKII alpha bind calmodulin with distinct binding affinities in live cell, which indicates that calcium signaling is fine-tuned by distinct activation patterns of CaM kinases. This method will enable investigation of cellular processes based on dynamic P-P interactions.

引用

页码：3412 / 3417

页数：6

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