kinesin-1 carpets |
standard tub, 10-20 μm taxol (after?) b polym. |
1-1.4 c
|
waves and curls upon pinning |
(1) |
standard tub, 50 μM taxol during & after polym.; high MT density (2.5 MTs/μm2) |
1-5 |
loops form through collision; loop duration frequently >5 min; strong increase in loops at high MT concentration; decreasing loop radius with increasing contour |
(2) |
rhodamine-tub, 10 μm taxol after polym.; exposing to air bubble or n-heptane |
1.1 (heptane), 1.8 (air) |
MTs become reversibly unstable in non-polar conditions: 50% of MTs form loops as long as close to air bubble; effect absolutely requires kinesins |
(3) |
rhodamine-tub, 10 μM taxol after polym. |
2.5-3.75 c
|
left-handed supertwist favours CCW rotation of loops; CCW rotation is preserved in spools |
(4) |
biotin-tub, 10 μm taxol after polym.; SA-linked |
1-12.6, mean 3.9 |
biotin-tub, 10 μm taxol after polym.; SA-linked |
1-5, mean 2.3 |
up to 25 μm long straight bundles; pinning of tip induces spools or fishtailing; occasional “unspooling” events |
(5) |
biotin-tub, 10 μm taxol after polym.; SA-linked; 1600, 870, 270 and 90 kinesins/μm2
|
ca. 2.4-4 |
highest spool density & lowest spool diameter @ highest kinesin density; pinning as main cause for spool formation |
(6) |
biotin-GTP-tub, 10 μm taxol after polym.; SA-linked |
5.7 (@ 10.8 μm length), 3 (@ 3,7 μm) |
spool diameters increase with MT length per condition; spool diameters: GMP-MTs (taxol) < GMPCPP-MTs (no taxol) < GMPCPP-MTs (taxol) |
(7) |
biotin-GMPCPP-tub, 10 μm taxol after polym.; SA-linked |
18.8 (@ 10.3 μm length), 5.8 (@ 3.4 μm) |
biotin-GMPCPP-tub, no taxol; SA-linked |
8.2 (@ 10 μm length), 4.3 (@ 3.4 μm) |
biotin-GTP-tub, 10 μm taxol (after?)b polym.; SA-linked |
3.2 μm (@ 6μm length) |
live imaging: pinning & collisions (simultaneous sticking) cause spool formation; spool formation is not activated by a Brownian ratchet type process |
(8) |
biotin-tub, 10 μm taxol after polym.; SA-linked; microfluidic device |
2.7 (pinning), 6.2 (collisions) |
live imaging: pinning & collisions (simultaneous sticking) cause spools of different diameters; pinning more frequent in flow cells than microfluidic device |
(9) |
biotin-tub, (taxol?)b polym.; SA-quantum dot-linked |
1.2, mean 3.4 |
left/right-handed super-twist: CCW/CW rotation; rings form intertwined wreath-like structures; tendency to disassemble involving MT breakage, kinesins pulling (blocked by AMP-PNP), counteracted by SA (enhanced by biotin) |
(10) |
biotin-tub, 10 μm taxol after polym.; SA-quantum dot-linked; patterned kinesin carpets |
1-5.3 and 3.1 |
smallest spool diameters on constrained carpets: 1-5.3 μm on 5 μm stripes, 3.1 μm on 2 μm wide squares |
(11) |
axonemal dynein carpet |
Cy3-tub, 10μM taxol |
straight |
forming vortices in mm range |
(12) |