Figure 1.
Pedigree diagram of family DP.
The proband is marked with an arrow. Black symbols indicate clinically affected family members, open symbols indicate unaffected, and crossed symbols indicate deceased persons. Family DP consisted of twelve members, of which only one participated in the study. Genetic analyses were performed for II:2 (the proband) and III:3 (proband's son).
Figure 2.
Pedigree diagram of family ZP.
Open diamond (IV:17). indicates sex unknown. Open triangle (V:4) indicates miscarriage. The other symbols indications as above. Family ZP consisted of 52 members, five of which participated in the study. Genetic analyses were performed for individuals III:7, IV:2, IV:4, IV:5, and IV:14.
Figure 3.
Pedigree diagram of family KP.
The symbols indications as above. Family KP consisted of 183 members, and eleven participated in the study. Genetic analyses were performed for III:1, III:11, IV:20, IV:25, IV:40, IV:44, V:7, V:8, V:9, V:43, V:44.
Figure 4.
Pedigree diagram of family RP.
The symbols indications as above. Family RP consisted of 27 members, three of which (IV:6, IV:8, V:7) participated in the previous study [18].
Table 1.
Haplotype analysis of the families with the DES mutations.
Figure 5.
Electron microscopy analysis of the longitudinal sections of probands's muscle biopsies.
Bars, 1 µm.
Figure 6.
Distribution of desmin in control muscle.
Transverse sections of muscle biopsy of a healthy subject were stained with anti-desmin, anti-M-cadherin and anti-slow myosin heavy chain (MHCs) antibodies, as indicated in the figure. Right panels, ∼2x magnification of the regions marked in left panels. These are 0.8-µm images of the center of transverses muscle section obtained with a Leica confocal microscope. Other details as described in Materials and Methods section. Bars, 50 µm.
Figure 7.
Distribution of desmin in muscle with A357_E359del mutation.
Transverse sections of the muscle biopsy of the patient were stained with anti-desmin, anti-M-cadherin and anti-slow myosin heavy chain antibodies, as indicated in the figure. Right panels, ∼2x magnification of the regions marked in left panels. These are 0.8-µm images of the center of transverses muscle section obtained with a Leica confocal microscope. Other details as described in Materials and Methods section. Bars, 50 µm.
Figure 8.
Distribution of desmin in muscle with Q348P mutation.
Transverse sections of the muscle biopsy of the patient were stained with anti-desmin, anti-M-cadherin, anti-α-actinin and anti-slow myosin heavy chain antibodies, as indicated in the figure. Right panels, ∼2x magnification of the regions marked in left panels. Arrows, centrally located nuclei; asterisk, an atrophic fiber. These are 0.8-µm images of the center of transverses muscle section obtained with a Leica confocal microscope. Details as described in Materials and Methods. Bars, 50 µm.
Figure 9.
Quantification of desmin content in muscle by immunoblotting.
Equal volumes of homogenates (A) and supernatants (B) from control (C1-C4), patient IV:2, ZP family (P1, A357_E359del mutation), and patient III:3, DP family (P2, Q348P mutation) muscle were subjected to SDS polyacrylamide gel electrophoresis, blotted on nitrocellulose membrane and probed with anti-desmin and anti-GAPDH antibodies, as described in Materials and Methods. Lower panels in A and B, densitometric analyses of desmin content in the examined muscles. For controls, the data are presented as mean ± SEM for n = 4.
Figure 10.
Histogram of helix bending angle at residue 348.
WT helix is naturally bent, about 175°, in a dimer coiled-coil conformation. For the Q348P mutation the maximum of the helix bending angle is nearly the same, about 172°, but a long tail of this plot with a local maximum at 158° indicates a high flexibility of helix at this residue in the mutant structure. The structure of A357-E359del also demonstrates a higher flexibility than WT.
Figure 11.
Alignment of WT (blue), point-mutated (yellow) and deletion (purple) structures of desmin dimer after a 30ns MD simulation.
Residue 348 is colored green in both helices.