Reverse actin sliding triggers strong myosin binding that moves tropomyosin.

Actin/myosin interactions in vertebrate striated muscles are believed to be regulated by the "steric blocking" mechanism whereby the binding of calcium to the troponin complex allows tropomyosin (TM) to change position on actin, acting as a molecular switch that blocks or allows myosin heads to interact with actin. Movement of TM during activation is initiated by interaction of Ca(2+) with troponin, then completed by further displacement by strong binding cross-bridges. We report x-ray evidence that TM in insect flight muscle (IFM) moves in a manner consistent with the steric blocking mechanism. We find that both isometric contraction, at high [Ca(2+)], and stretch activation, at lower [Ca(2+)], develop similarly high x-ray intensities on the IFM fourth actin layer line because of TM movement, coinciding with x-ray signals of strong-binding cross-bridge attachment to helically favored "actin target zones." Vanadate (Vi), a phosphate analog that inhibits active cross-bridge cycling, abolishes all active force in IFM, allowing high [Ca(2+)] to elicit initial TM movement without cross-bridge attachment or other changes from relaxed structure. However, when stretched in high [Ca(2+)], Vi-"paralyzed" fibers produce force substantially above passive response at pCa approximately 9, concurrent with full conversion from resting to active x-ray pattern, including x-ray signals of cross-bridge strong-binding and TM movement. This argues that myosin heads can be recruited as strong-binding "brakes" by backward-sliding, calcium-activated thin filaments, and are as effective in moving TM as actively force-producing cross-bridges. Such recruitment of myosin as brakes may be the major mechanism resisting extension during lengthening contractions.

Atomic force microscope study of presmectic modulation in the nematic and isotropic phases of the liquid crystal octylcyanobiphenyl using piezoresistive force detection.

Using a temperature controlled atomic force microscope (AFM), we have studied surface induced pre-smectic order in the nematic and isotropic phases of 4-cyano- 4'-n -octylbiphenyl. A modified AFM head with piezoresitive cantilevers has been used to measure the structural force between a flat BK7 glass plate and a 10 microm glass sphere, both being treated to induce homeotropic alignment of the confined liquid crystal layer in between. We have observed surface-induced presmectic force not only in the isotropic, but also in the nematic phase. We have measured the temperature dependencies of the presmectic force, the smectic correlation length xi and the smectic order parameter psi at the surface. The correlation length xi(T) shows a power-law temperature dependence with a critical exponent of nu=0.67 +/- 0.03 and the bare correlation length of xi(0) = (0.39 +/- 0.08) nm, in good agreement with x-ray data. The smectic density at the surface is psi(2)(S) =0.4 in the nematic phase and decreases in the isotropic phase.