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Supplementary Figure 1
Longitudinal view of reconstruction density seen in projection,
demonstrating arrowheads (cf. Fig. 1a of paper)
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Supplementary Figure 2
Longitudinal surface view of reconstruction, showing how the transverse
view in Fig. 2a of paper is obtained (see Supplementary Movies S2 and
S3).
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Supplementary Figure S3
Transverse view of reconstruction seen in projection, demonstrating
high density of subfilaments and low density of filament core (cf. Fig.
2b of paper).
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Supplementary Movie 1
Movie of longitudinal view of reconstruction, including three complete
crowns of heads, rotated around longitudinal axis (bare zone direction
at top). Compare Fig. 1c of paper. Each crown contains four J-motifs
separated by 90 degrees. The filament rotates 90 degrees, revealing all
unique views, owing to the 4-fold rotational symmetry of the filament.
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Movie (3.1MB)
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Supplementary Movie 2
Movie showing how longitudinal view of reconstruction transforms into
transverse view seen in Fig. 2a of paper.
This movie helps interpretation of Fig. 2a
of the paper. It first shows a longitudinal view of the reconstruction,
including only a single crown of heads (see Supplementary Fig. 2). The
crown rotates 90 degrees, then becomes truncated to include only a 14.5
nm repeat (slightly shorter than a crown, owing to motif overlap). This
segment then rotates 90 degrees towards the viewer to show a transverse
view from the direction of the bare zone, as seen in Fig. 2a of the
paper. The motif originally located at the front of the reconstruction
is now seen to be located at the bottom, whereas the motif originally
hidden at the back of the reconstruction is now at the top. The segment
now rotates 90 degrees around the filament axis to produce an identical
view, illustrating the 4-fold rotational symmetry. Finally, the segment
is rotated in three 30 degree steps to simulate the views obtained when
moving by 14.5 nm steps along the filament axis away from the bare
zone: the helical symmetry of the tarantula filament is such that each
segment is rotated 30 degrees relative to the previous one (see
Supplementary Movie 3).
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Supplementary Movie 3
This movie shows how portions of different crowns contribute to the
transverse view of the reconstruction containing a single 14.5 nm
repeat seen in Fig. 2a of paper and Supplementary Movie 2):
The 14.5 nm thick segment is initially
positioned and oriented as in Fig. 2a, and then progressively moves
away from the bare zone through two more repeats. The filament appears
to rotate by 30 degrees for each 14.5 nm of movement along the filament
axis (see Supplementary Movie 2). The initial and final positions of
the segments are shown in Supplementary Fig. 2.
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Movie (1.9MB)
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Supplementary Movie 4
This movie provides a three-dimensional perspective on the fitting of
the heads of the modified smooth muscle HMM atomic structure (PDB1i843)
into the J-motif of the filament reconstruction (cf. Fig. 3a of paper):
The filament is oriented with the bare zone
at the top. The view moves from one side of the motif to the other
around a vertical axis (cf. Fig. 1 of paper and Supplementary Movie 1).
A rod-like region of density can be seen connecting the bottom of the
"J" structure to the underlying surface of the filament core (red
asterisk in Fig. 3a of paper). This is the proposed location of the
initial (N-terminal) portion of the S2 fragment of the myosin tail. A
small volume of unfilled density in the light chain domains could
accommodate the putative 50% larger volume of the tarantula regulatory
light chain (Ref. 26) compared with the vertebrate RLC used in the
fitting. Note: while the new model differs from the "splayed heads"
structures proposed previously (see Fig. 3 legend of paper), the
current and earlier models agree in having interactions between myosin
heads that are on the same helical track but in adjacent crowns (see
Fig. 5 of paper).
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Supplementary Movie 5
This movie shows different views of the space filling atomic model of
PDB 1i84 after modification to fit the "J" motif of the reconstruction
(cf. Figs 3b and 5a of paper). The bare zone direction is towards the
top. The model rotates around a vertical axis through 360 degrees to
show views from all sides.
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Movie (8MB)
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Supplementary Movie 6
Movie showing how S2 fits into the reconstruction (cf. Fig. 4 of paper):
The initial view shows the J-motif (bare
zone at top) fitted with the two heads (Fig. 3a of paper), together
with the N-terminal portion of the associated S2 (red; cf. Fig. 5b of
paper). The view moves through 90 degrees to expose the S2 and its
sloping path towards the subfilaments, and then rotates to produce a
horizontal orientation, with bare zone towards the right (cf. Fig. 4a
of paper). The view now rotates around a vertical axis, reaching an
orientation similar to that in Fig. 4b of the paper after 90 degrees,
then continuing to complete a 360 degree rotation, showing the two
heads and the S2 from all sides in this horizontal plane
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Movie (7.3MB)
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