Back to Search
Start Over
Multiple Sites in αB-Crystallin Modulate Its Interactions with Desmin Filaments Assembled In Vitro
- Source :
- PLoS ONE, PLoS ONE, 2011, Vol.6(11), pp.e25859 [Peer Reviewed Journal], PLoS ONE, Vol 6, Iss 11, p e25859 (2011)
- Publication Year :
- 2011
- Publisher :
- Public Library of Science (PLoS), 2011.
-
Abstract
- The β3- and β8-strands and C-terminal residues 155-165 of αB-crystallin were identified by pin arrays as interaction sites for various client proteins including the intermediate filament protein desmin. Here we present data using 5 well-characterised αB-crystallin protein constructs with substituted β3- and β8-strands and with the C-terminal residues 155-165 deleted to demonstrate the importance of these sequences to the interaction of αB-crystallin with desmin filaments. We used electron microscopy of negatively stained samples to visualize increased interactions followed by sedimentation assays to quantify our observations. A low-speed sedimentation assay measured the ability of αB-crystallin to prevent the self-association of desmin filaments. A high-speed sedimentation assay measured αB-crystallin cosedimentation with desmin filaments. Swapping the β8-strand of αB-crystallin or deleting residues 155-165 increased the cosedimentation of αB-crystallin with desmin filaments, but this coincided with increased filament-filament interactions. In contrast, substitution of the β3-strand with the equivalent αA-crystallin sequences improved the ability of αB-crystallin to prevent desmin filament-filament interactions with no significant change in its cosedimentation properties. These data suggest that all three sequences (β3-strand, β8-strand and C-terminal residues 155-165) contribute to the interaction of αB-crystallin with desmin filaments. The data also suggest that the cosedimentation of αB-crystallin with desmin filaments does not necessarily correlate with preventing desmin filament-filament interactions. This important observation is relevant not only to the formation of the protein aggregates that contain both desmin and αB-crystallin and typify desmin related myopathies, but also to the interaction of αB-crystallin with other filamentous protein polymers.
- Subjects :
- lcsh:Medicine
macromolecular substances
Plasma protein binding
Protein aggregation
Biology
Cardiovascular
Biochemistry
Desmin
Protein–protein interaction
03 medical and health sciences
0302 clinical medicine
Microscopy, Electron, Transmission
Crystallin
Molecular Cell Biology
Humans
Intermediate Filament Protein
Binding site
lcsh:Science
Protein Interactions
Cytoskeleton
Cellular Stress Responses
030304 developmental biology
0303 health sciences
Binding Sites
Multidisciplinary
lcsh:R
Proteins
Crystallins
Molecular biology
Recombinant Proteins
Cellular Structures
eye diseases
Chaperone Proteins
Cytoskeletal Proteins
Mutagenesis, Site-Directed
Biophysics
Medicine
lcsh:Q
Structural Proteins
sense organs
Cardiomyopathies
030217 neurology & neurosurgery
Research Article
Protein Binding
Subjects
Details
- ISSN :
- 19326203
- Volume :
- 6
- Database :
- OpenAIRE
- Journal :
- PLoS ONE
- Accession number :
- edsair.doi.dedup.....c3e4d11f50a4f90f36b38e27a60cbcca