Dataset Information

The N-cadherin interactome in primary cardiomyocytes as defined by quantitative proximity proteomics

ABSTRACT: Low levels of Cdh2-BioID2 were expressed in mouse primary neonatal cardiomyocytes using an adenoviral expression system. Infected cells were treated with biotin for 24 hours before lysis. Biotinylated proteins were precipitated from lysates using streptavidin beads. Protein samples were in-gel trypsin digested and the resulting tryptic peptides were analyzed on an LTQ Orbitrap Velos.

INSTRUMENT(S): LTQ Orbitrap Velos

ORGANISM(S): Mus Musculus (ncbitaxon:10090)

SUBMITTER: Adam V. Kwiatkowski

PROVIDER: MSV000083362 | MassIVE | Thu Jan 24 18:34:00 GMT 2019

SECONDARY ACCESSION(S): PXD012496

REPOSITORIES: MassIVE

ACCESS DATA

Dataset's files

Source:

			Action	DRS
		Other

Items per page:

1 - 1 of 1

Publications

The N-cadherin interactome in primary cardiomyocytes as defined using quantitative proximity proteomics.

Li Yang Y Merkel Chelsea D CD Zeng Xuemei X Heier Jonathon A JA Cantrell Pamela S PS Sun Mai M Stolz Donna B DB Watkins Simon C SC Yates Nathan A NA Kwiatkowski Adam V AV

Journal of cell science 20190211 3

The junctional complexes that couple cardiomyocytes must transmit the mechanical forces of contraction while maintaining adhesive homeostasis. The adherens junction (AJ) connects the actomyosin networks of neighboring cardiomyocytes and is required for proper heart function. Yet little is known about the molecular composition of the cardiomyocyte AJ or how it is organized to function under mechanical load. Here, we define the architecture, dynamics and proteome of the cardiomyocyte AJ. Mouse neo ...[more]

PMID: 30630894

Similar Datasets

Project description:Classical methods of investigating protein-protein interactions (PPIs) are generally performed in non-living systems, yet in recent years new technologies utilizing proximity labeling (PL) have given researchers the tools to explore proximal PPIs in living systems. PL has distinct advantages over traditional protein interactome studies, such as the ability to identify weak and transient interactions in vitro and in vivo. Most PL studies are performed on targets within or on the cell membrane. We have adapted the original PL method to investigate PPIs within the extracellular compartment, using both BioID2 and TurboID, that we term extracellular PL (ePL). To demonstrate the utility of this modified technique, we investigate the interactome of the widely expressed matrisome protein Tissue inhibitors of metalloproteinases 2 (TIMP2). Tissue inhibitors of metalloproteinases (TIMPs) are a family of multi-functional proteins that were initially defined by their ability to inhibit the enzymatic activity of metalloproteinases (MPs), the major mediators of extracellular matrix (ECM) breakdown and turnover. TIMP2 exhibits a broad expression profile and is often abundant in both normal and diseased tissues. Understanding the functional transformation of matrisome regulators, like TIMP2, during the evolution of tissue microenvironments associated with disease progression is essential for the development of ECM targeted therapeutics. Using carboxyl- and amino-terminal fusion proteins of TIMP2 with BioID2 and TurboID, we describe the TIMP2 proximal interactome. We also illustrate how the TIMP2 interactome changes in the presence of different stimuli, in different cell types, in unique culture conditions (2D vs 3D), and with different reaction kinetics (BioID2 vs. TurboID); demonstrating the power of this technique versus classical PPI methods. We propose that the screening of matrisome targets in disease models using ePL will reveal new therapeutic targets for further comprehensive studies.