Synemin: Molecular Features and the Use of Proximity Ligation Assay to Study Its Interactions.

Abstract

Synemin has three splice variants (α, β, and L) with identical head and rod domains but with tail domains of varying size. α- and β-Synemin are larger than most intermediate filament proteins (1565 and 1253 amino acids, respectively) but L-synemin is shorter (339 amino acids). Synemin isoforms do not self-assemble into filaments but can copolymerize with vimentin and desmin. Synemin is present in all muscle cell types, in a few neural cell types, and in various other nonepithelial cell types. Synemin expression is regulated, sometimes in an isoform-specific manner, during development of the nervous system, in brain and breast cancer cells and during injuries to the brain and liver. Mice-lacking synemin develop a myopathic phenotype, possibly due to synemin role in linking desmin filaments to costameres and sarcomeres. Synemin may play this role through its demonstrated binding to costameric and sarcolemmal proteins, such as α-actinin, vinculin, and members of the dystroglycan complex. In astrocytoma cells, synemin regulates proliferation by interacting with PP2A to modulate Akt phosphorylation status. Methods to identify synemin binding partners are central to understand the roles of this protein in diverse cell types. Here, we describe how to use proximal ligation assays (PLA) for this purpose. PLA complement biochemical methods such as immunoprecipitation by relying on the use of antibodies conjugated to oligonucleotide probes to visualize by fluorescence microscopy protein-protein interactions in cells and tissues.

• Authors: Madhumita Paul, Omar Skalli
• Journal: Methods in enzymology (Methods Enzymol) Vol. 568 Pg. 537-55 ( 2016) ISSN: 1557-7988 [Electronic] United States
• PMID: 26795483 (Publication Type: Journal Article)
• Copyright: Copyright © 2016 Elsevier Inc. All rights reserved.

Chemical References

• Dystrophin-Associated Proteins
• Intermediate Filament Proteins
• desmuslin
• dystrobrevin
• Protein Phosphatase 2

Topics

• Animals
• Astrocytes (metabolism)
• Dystrophin-Associated Proteins (metabolism)
• Humans
• Intermediate Filament Proteins (metabolism)
• Neurons (metabolism)
• Protein Phosphatase 2 (metabolism)