The Nonlinear Structure of the Desmoplakin Plakin Domain and the Effects of Cardiomyopathy-Linked Mutations

Al-Jassar, Caezar; Knowles, Timothy J.; Jeeves, Mark; Kami, Keiichiro; Behr, Elijah R.; Bikker, Hennie; Overduin, Michael; Chidgey, Martyn

doi:10.1016/j.jmb.2011.06.047

Cited by 29 publications

(47 citation statements)

References 47 publications

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“…The N-terminal plakin domain of desmoplakin has an 'L' conformation consisting of two perpendicular lobes separated by a flexible elbow, with a flexible linker connecting the N-terminal to the rod domain (AlJassar et al, 2011; Grum et al, 1999). Our data supports previously proposed models in which a conformational change within the plakin domain elbow or the flexible linker allows for motion of the rod and C-terminal domains within the plaque, whereas the N-terminal domain is stationary (Al-Jassar et al, 2011). PKP-1 has also been shown to interfere with plakoglobin-desmoplakin binding (Bornslaeger et al, 2001) and thus it is possible that the proposed conformational change also reflects a change in binding partner.…”

Section: Reorganization Of Plaque Proteins In Pkp-1-mediated Hyperadhsupporting

confidence: 90%

Molecular organization of the desmosome as revealed by direct stochastic optical reconstruction microscopy

Stahley

Bartle

Atkinson

et al. 2016

Journal of Cell Science

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Desmosomes are macromolecular junctions responsible for providing strong cell-cell adhesion. Because of their size and molecular complexity, the precise ultrastructural organization of desmosomes is challenging to study. Here, we used direct stochastic optical reconstruction microscopy (dSTORM) to resolve individual plaque pairs for inner and outer dense plaque proteins. Analysis methods based on desmosomal mirror symmetry were developed to measure plaque-to-plaque distances and create an integrated map. We quantified the organization of desmoglein 3, plakoglobin and desmoplakin (N-terminal, rod and C-terminal domains) in primary human keratinocytes. Longer desmosome lengths correlated with increasing plaque-to-plaque distance, suggesting that desmoplakin is arranged with its long axis at an angle within the plaque. We next examined whether plaque organization changed in different adhesive states. Plaque-to-plaque distance for the desmoplakin rod and C-terminal domains decreased in PKP-1-mediated hyperadhesive desmosomes, suggesting that protein reorganization correlates with function. Finally, in human epidermis we found a difference in plaque-to-plaque distance for the desmoplakin Cterminal domain, but not the desmoplakin rod domain or plakoglobin, between basal and suprabasal cells. Our data reveal the molecular organization of desmosomes in cultured keratinocytes and skin as defined by dSTORM.

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Section: Reorganization Of Plaque Proteins In Pkp-1-mediated Hyperadhsupporting

confidence: 90%

Molecular organization of the desmosome as revealed by direct stochastic optical reconstruction microscopy

Stahley

Bartle

Atkinson

et al. 2016

Journal of Cell Science

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“…The conserved local environment of plectin Lys-1156 and desmoplakin Arg-808 suggest that R808C might alter the helical bundle locally and expose a hydrophobic patch on the surface of the SR8. This is in agreement with the destabilizing effect of R808C when introduced in an SR7-SR8 construct (32). Finally, desmoplakin Ile-870 corresponds to Val-1218 of plectin, which is part of the hydrophobic core of the SR8; hence, the substitution I870M is likely to perturb the stability of this SR. Other missense mutations within the SR7-SR9 region of desmoplakin have been observed in healthy individuals.…”

supporting

confidence: 80%

“…In SR7-SR9 of desmoplakin, mutations N661I, Y787C, R808C, and R808H, have been linked to arrhythmogenic right ventricular dysplasia/cardiomyopathy (ARVD/C) (32)(33)(34), and the mutation I870M has been observed in a patient with dilated cardiomyopathy (35). Owing to the high similarity between the SR7-SR9 of plectin and desmoplakin, we analyzed the possible structural effects of these mutations in desmoplakin using the high resolution structure of plectin as a template (Fig 12).…”

mentioning

confidence: 99%

The Structure of the Plakin Domain of Plectin Reveals an Extended Rod-like Shape

Ortega

Manso

Buey

et al. 2016

Journal of Biological Chemistry

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Plakins are large multi-domain proteins that interconnect cytoskeletal structures. Plectin is a prototypical plakin that tethers intermediate filaments to membrane-associated complexes. Most plakins contain a plakin domain formed by up to nine spectrin repeats (SR1-SR9) and an SH3 domain. The plakin domains of plectin and other plakins harbor binding sites for junctional proteins. We have combined x-ray crystallography with small angle x-ray scattering (SAXS) to elucidate the structure of the plakin domain of plectin, extending our previous analysis of the SR1 to SR5 region. Two crystal structures of the SR5-SR6 region allowed us to characterize its uniquely wide inter-repeat conformational variability. We also report the crystal structures of the SR7-SR8 region, refined to 1.8 Å, and the SR7-SR9 at lower resolution. The SR7-SR9 region, which is conserved in all other plakin domains, forms a rigid segment stabilized by uniquely extensive inter-repeat contacts mediated by unusually long helices in SR8 and SR9. Using SAXS we show that in solution the SR3-SR6 and SR7-SR9 regions are rod-like segments and that SR3-SR9 of plectin has an extended shape with a small central kink. Other plakins, such as bullous pemphigoid antigen 1 and microtubule and actin cross-linking factor 1, are likely to have similar extended plakin domains. In contrast, desmoplakin has a two-segment structure with a central flexible hinge. The continuous versus segmented structures of the plakin domains of plectin and desmoplakin give insight into how different plakins might respond to tension and transmit mechanical signals.Plakins are a family of very large proteins that interconnect and organize the intermediate filaments (IF), 4 microtubules, and microfilaments of the cytoskeleton and tether them to membrane-associated structures (1, 2). So far, seven plakins have been described in mammals; these are plectin, bullous pemphigoid antigen 1 (BPAG1), desmoplakin, microtubule and actin cross-linking factor 1 (MACF1, also known as ACF7), envoplakin, periplakin, and epiplakin. Invertebrates have a more reduced plakin repertoire; for example Caenorhabditis elegans and Drosophila melanogaster each have a single plakin gene encoding VAB-10 and Shot (also known as Short Stop or kakapo), respectively. Most of the plakin genes produce multiple isoforms that increase the structural and functional versatility of these proteins.Plectin is expressed in a large variety of cell types in which it acts as a highly polyvalent cytolinker that contributes to cell adhesion and the organization of the cytoskeleton. Plectin cross-links IFs to microtubules and actin filaments and mediates the attachment of IFs to cell-cell and cell-matrix junctional complexes such as hemidesmosomes, desmosomes, Z-lines, and focal contacts. Plectin also connects IFs to organelles such as the nucleus and mitochondria (3). Defects in the PLEC gene cause various forms of the blistering disease epidermolysis bullosa simplex (EBS), which may occur only with skin fragility, as found in EBS Og...

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“…A splice variant eliminates part of the central rod domain to produce DPII (Green et al, 1990). DPNT consists of an amino-terminal basic region (residues 1-170) that contains the binding site for plakoglobin (Kowalczyk et al, 1997;Yang et al, 2006) followed by a plakin domain, which binds Dsg1 and PKP1 (Al-Jassar et al, 2011;Kami, Chidgey, Dafforn, & Overduin, 2009;Kowalczyk et al, 1999). The plakin domains of plakin family members, which include DP, bullous pemphigoid antigen 1 (BPAG1), plectin, envoplakin, and periplakin, contain variable numbers of spectrin-like repeats, as first shown by sequence analysis enabled by the crystal structures of di-repeat fragments of plectin and BPAG1 ( Jefferson, Ciatto, Shapiro, & Liem, 2007;Sonnenberg, Rojas, & de Pereda, 2007).…”

Section: Introductionmentioning

confidence: 99%

“…1). Amino acid sequence analysis and protease sensitivity (see later) suggest that there is a flexible linker between DP SR6 and SR7, and a single SH3 domain is inserted in the middle of SR5 (Al-Jassar, Bernado, Chidgey, & Overduin, 2013;Al-Jassar et al, 2011;Choi & Weis, 2011;Jefferson et al, 2007;.…”

Section: Introductionmentioning

confidence: 99%

Purification and Structural Analysis of Desmoplakin

Weis

2016

Methods in Enzymology

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The Nonlinear Structure of the Desmoplakin Plakin Domain and the Effects of Cardiomyopathy-Linked Mutations

Cited by 29 publications

References 47 publications

Molecular organization of the desmosome as revealed by direct stochastic optical reconstruction microscopy

Molecular organization of the desmosome as revealed by direct stochastic optical reconstruction microscopy

The Structure of the Plakin Domain of Plectin Reveals an Extended Rod-like Shape

Purification and Structural Analysis of Desmoplakin

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