2. Academic Validation
  3. Spatiotemporal profiling of cytosolic signaling complexes in living cells by selective proximity proteomics

Spatiotemporal profiling of cytosolic signaling complexes in living cells by selective proximity proteomics

  • Nat Commun. 2021 Jan 4;12(1):71. doi: 10.1038/s41467-020-20367-x.
Mi Ke 1,  Xiao Yuan 1,  An He 1,  Peiyuan Yu 1,  Wendong Chen 1,  Yu Shi 2,  Tony Hunter 2,  Peng Zou 3,  Ruijun Tian 4 5 6
Affiliations

Affiliations

  • 1 Department of Chemistry, School of Science, Southern University of Science and Technology, Shenzhen, China.
  • 2 Molecular and Cell Biology Laboratory, Salk Institute for Biological Studies, La Jolla, CA, USA.
  • 3 College of Chemistry and Molecular Engineering, Peking University, Beijing, China.
  • 4 Department of Chemistry, School of Science, Southern University of Science and Technology, Shenzhen, China. tianrj@sustech.edu.cn.
  • 5 Guangdong Provincial Key Laboratory of Cell Microenvironment and Disease Research, Southern University of Science and Technology, Shenzhen, China. tianrj@sustech.edu.cn.
  • 6 Shenzhen Grubbs Institute, Southern University of Science and Technology, 518055, Shenzhen, China. tianrj@sustech.edu.cn.
PMID: 33397984 DOI: 10.1038/s41467-020-20367-x
Abstract

Signaling complexes are often organized in a spatiotemporal manner and on a minute timescale. Proximity labeling based on engineered ascorbate peroxidase APEX2 pioneered in situ capture of spatiotemporal membrane protein complexes in living cells, but its application to cytosolic proteins remains limited due to the high labeling background. Here, we develop proximity labeling probes with increased labeling selectivity. These probes, in combination with label-free quantitative proteomics, allow exploring cytosolic protein assemblies such as phosphotyrosine-mediated protein complexes formed in response to minute-scale EGF stimulation. As proof-of-concept, we systematically profile the spatiotemporal interactome of the EGFR signaling component STS1. For STS1 core complexes, our proximity proteomics approach shows comparable performance to affinity purification-mass spectrometry-based temporal interactome profiling, while also capturing additional-especially endosomally-located-protein complexes. In summary, we provide a generic approach for exploring the interactome of mobile cytosolic proteins in living cells at a temporal resolution of minutes.

Figures
Products
嗨！很高兴为您提供帮助！

尊敬的 MCE 客户您好，
请您选择所在区域，我们将转接对应客服为您服务！

热门区域

A

B

C

F

G

H

J

L

N

Q

S

T

X

Y

Z

热 A B C F G H J L N Q S T X Y Z