Stabilization of vimentin coil2 fragment via an engineered disulfide

Chernyatina, A.A.; Strelkov, S.V.

doi:10.1016/j.jsb.2011.11.014

Cited by 28 publications

(33 citation statements)

References 39 publications

(57 reference statements)

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Together with our recent results on the complete coil-2 structure (19,20), the data presented here allow us to reconstruct the vimentin rod in its entire length (Fig. 4A).…”

Section: Discussionsupporting

confidence: 65%

“…3A. Similarly, the coil-2 structure (blue) is a superposition of previously established fragment structures (19,42). The linker L12 for which no crystallographic data are available yet is shown schematically in pink.…”

Section: Discussionmentioning

confidence: 99%

“…We are focusing on X-ray crystallographic studies of human vimentin as a "model" IF protein, using a "divide-and-conquer" approach based on the crystallization of overlapping short fragments (17). These efforts resulted in the atomic structure determination of the vimentin coil 1A [Protein Data Bank (PDB) entry 1GK7] (18) and of several overlapping fragments covering the complete coil 2 [PDB entries 1GK6 (18), 3KLT (19), and 3TRT (20)]. An important conclusion of these studies was that coil 2 forms a contiguous CC structure-i.e., the linker L2 does not exist.…”

mentioning

confidence: 99%

See 2 more Smart Citations

Atomic structure of the vimentin central α-helical domain and its implications for intermediate filament assembly

Chernyatina

Nicolet

Aebi

et al. 2012

Proc. Natl. Acad. Sci. U.S.A.

157

164

View full text Add to dashboard Cite

Together with actin filaments and microtubules, intermediate filaments (IFs) are the basic cytoskeletal components of metazoan cells. Over 80 human diseases have been linked to mutations in various IF proteins to date. However, the filament structure is far from being resolved at the atomic level, which hampers rational understanding of IF pathologies. The elementary building block of all IF proteins is a dimer consisting of an α-helical coiled-coil (CC) "rod" domain flanked by the flexible head and tail domains. Here we present three crystal structures of overlapping human vimentin fragments that comprise the first half of its rod domain. Given the previously solved fragments, a nearly complete atomic structure of the vimentin rod has become available. It consists of three α-helical segments (coils 1A, 1B, and 2) interconnected by linkers (L1 and L12). Most of the CC structure has a left-handed twist with heptad repeats, but both coil 1B and coil 2 also exhibit untwisted, parallel stretches with hendecad repeats. In the crystal structure, linker L1 was found to be α-helical without being involved in the CC formation. The available data allow us to construct an atomic model of the antiparallel tetramer representing the second level of vimentin assembly. Although the presence of the nonhelical head domains is essential for proper tetramer stabilization, the precise alignment of the dimers forming the tetramer appears to depend on the complementarity of their surface charge distribution patterns, while the structural plasticity of linker L1 and coil 1A plays a role in the subsequent IF assembly process.X-ray crystallography | cytoskeleton | coiled coil

show abstract

“…Together with our recent results on the complete coil-2 structure (19,20), the data presented here allow us to reconstruct the vimentin rod in its entire length (Fig. 4A).…”

Section: Discussionsupporting

confidence: 65%

Section: Discussionmentioning

confidence: 99%

mentioning

confidence: 99%

See 1 more Smart Citation

Atomic structure of the vimentin central α-helical domain and its implications for intermediate filament assembly

Chernyatina

Nicolet

Aebi

et al. 2012

Proc. Natl. Acad. Sci. U.S.A.

157

164

View full text Add to dashboard Cite

show abstract

“…Modelling was performed using the crystal structures of vimentin (PDB code 3TRT) and Maf-G transcription factor (PDB code 3A5T) as the templates for the coil and linker regions, respectively [29], [30]. According to the alignment the residues were mutated with the Whatif molecular modelling tool.…”

Section: Methodsmentioning

confidence: 99%

Intestinal Cell Barrier Function In Vitro Is Severely Compromised by Keratin 8 and 18 Mutations Identified in Patients with Inflammatory Bowel Disease

et al. 2014

View full text Add to dashboard Cite

Keratin 8 and 18 (K8/K18) mutations have been implicated in the aetiology of certain pathogenic processes of the liver and pancreas. While some K8 mutations (K8 G62C, K8 K464N) are also presumed susceptibility factors for inflammatory bowel disease (IBD), the only K18 mutation (K18 S230T) discovered so far in an IBD patient is thought to be a polymorphism. The aim of our study was to demonstrate that these mutations might also directly affect intestinal cell barrier function. Cell monolayers of genetically engineered human colonocytes expressing these mutations were tested for permeability, growth rate and resistance to heat-stress. We also calculated the change in dissociation constant (Kd, measure of affinity) each of these mutations introduces into the keratin protein, and present the first model of a keratin dimer L12 region with in silico clues to how the K18 S230T mutation may affect keratin function. Physiologically, these mutations cause up to 30% increase in paracellular permeability in vitro. Heat-stress induces little keratin clumping but instead cell monolayers peel off the surface suggesting a problem with cell junctions. K18 S230T has pronounced pathological effects in vitro marked by high Kd, low growth rate and increased permeability. The latter may be due to the altered distribution of tight junction components claudin-4 and ZO-1. This is the first time intestinal cells have been suggested also functionally impaired by K8/K18 mutations. Although an in vitro colonocyte model system does not completely mimic the epithelial lining of the intestine, nevertheless the data suggest that K8/K18 mutations may be also able to produce a phenotype in vivo.

show abstract

“…Crystallization studies of different vimentin fragments indicate that the N-terminal residues 306-340 alone either do not fold or form very labile CCs at most (6,23). Indeed, our region I corresponds to this stretch of amino acids and exhibits very low stability, with the first eight N-terminal amino acids presumably being unstructured, as explained above.…”

Section: Discussion Entropic Cost For Seed Formation and Exchanged Simentioning

confidence: 81%

Sequence-resolved free energy profiles of stress-bearing vimentin intermediate filaments

Ramm

Stigler

Hinczewski

et al. 2014

Proc. Natl. Acad. Sci. U.S.A.

View full text Add to dashboard Cite

Intermediate filaments (IFs) are key to the mechanical strength of metazoan cells. Their basic building blocks are dimeric coiled coils mediating hierarchical assembly of the full-length filaments. Here we use single-molecule force spectroscopy by optical tweezers to assess the folding and stability of coil 2B of the model IF protein vimentin. The coiled coil was unzipped from its N and C termini. When pulling from the C terminus, we observed that the coiled coil was resistant to force owing to the high stability of the C-terminal region. Pulling from the N terminus revealed that the N-terminal half is considerably less stable. The mechanical pulling assay is a unique tool to study and control seed formation and structure propagation of the coiled coil. We then used rigorous theory-based deconvolution for a model-free extraction of the energy landscape and local stability profiles. The data obtained from the two distinct pulling directions complement each other and reveal a tripartite stability of the coiled coil: a labile N-terminal half, followed by a medium stability section and a highly stable region at the far C-terminal end. The different stability regions provide important insight into the mechanics of IF assembly.protein folding | Brownian dynamics simulation | trigger sequence

show abstract

Stabilization of vimentin coil2 fragment via an engineered disulfide

Cited by 28 publications

References 39 publications

Atomic structure of the vimentin central α-helical domain and its implications for intermediate filament assembly

Atomic structure of the vimentin central α-helical domain and its implications for intermediate filament assembly

Intestinal Cell Barrier Function In Vitro Is Severely Compromised by Keratin 8 and 18 Mutations Identified in Patients with Inflammatory Bowel Disease

Sequence-resolved free energy profiles of stress-bearing vimentin intermediate filaments

Contact Info

Product

Resources

About