Viscoelastic properties of f-actin, microtubules, f-actin/alpha-actinin, and f-actin/hexokinase determined in microliter volumes with a novel nondestructive method.
A nondestructive method to determine viscoelastic properties of gels and fluids involves an oscillating glass fiber serving as a sensor for the viscosity of the surrounding fluid. Extremely small displacements (typically 1-100 nm) are caused by the glass rod oscillating at its resonance frequency. These displacements are analyzed using a phase-sensitive acoustic microscope. Alterations of the elastic modulus of a fluid or gel change the propagation speed of a longitudinal acoustic wave. The system allows to study quantities as small as 10 microliters with temporal resolution >1 Hz. For 2-100 microM f-actin gels a final viscosity of 1.3-9.4 mPa s and a final elastic modulus of 2.229-2.254 GPa (corresponding to 1493-1501 m/s sound velocity) have been determined. For 10- to 100-microM microtubule gels (native, without stabilization by taxol), a final viscosity of 1.5-124 mPa s and a final elastic modulus of 2.288-2. 547 GPa (approximately 1513-1596 m/s) have been determined. During polymerization the sound velocity in low-concentration actin solutions increased up to +1.3 m/s (approximately 1.69 kPa) and decreased up to -7 m/s (approximately 49 kPa) at high actin concentrations. On polymerization of tubulin a concentration-dependent decrease of sound velocity was observed, too (+48 to -12 m/s approximately 2.3-0.1 MPa, for 10- to 100-microM tubulin). This decrease was interpreted by a nematic phase transition of the actin filaments and microtubules with increasing concentration. 2 mM ATP (when compared to 0.2 mM ATP) increased polymerization rate, final viscosity and elastic modulus of f-actin (17 microM). The actin-binding glycolytic enzyme hexokinase also accelerated the polymerization rate and final viscosity but elastic modulus (2.26 GPa) was less than for f-actin polymerized in presence of 0.2 mM ATP (2.28 GPa).
Wagner
O
O
Zoologisches, Physikalisches Institut der Johann Wolfgang Goethe Universität-Frankfurt/M, Frankfurt, Germany.
Zinke
J
J
Dancker
P
P
Grill
W
W
Bereiter-Hahn
J
J
D000162:Acoustics; D000185:Actinin / Q000737:chemistry*; D000199:Actins / Q000737:chemistry* / Q000648:ultrastructure; D000818:Animals; D055592:Biophysical Phenomena; D001703:Biophysics; D004548:Elasticity; D005782:Gels; D006593:Hexokinase / Q000737:chemistry*; D008854:Microscopy, Electron; D008870:Microtubules / Q000737:chemistry*; D009991:Oscillometry / Q000295:instrumentation; D010590:Phalloidine / Q000737:chemistry; D011817:Rabbits; D012212:Rheology / Q000295:instrumentation; D014404:Tubulin / Q000737:chemistry; D014783:Viscosity
D000199:Actins; D005782:Gels; D014404:Tubulin; D000185:Actinin; D010590:Phalloidine; D006593:Hexokinase