Buckling of a single microtubule by optical trapping forces: direct measurement of microtubule rigidity.

As major determinants of cell shape and polarity, microtubules are required to have suitable rigidity. However, our knowledge of the mechanical properties of microtubules is far from satisfactory. We report here a new method of measuring the flexural rigidity of a single microtubule by direct buckling using the optical trapping technique. Microtubule buckling was induced by applying a small longitudinal compressing force through an optically trapped microsphere that was firmly attached to the microtubule. Three ways of estimating the flexural rigidity of a continuous slender rod, one from the observed critical load of buckling and two from deflected lengths and angles of bending, yielded values which agreed well when applied to the analysis of buckling microtubules. Unexpectedly, we found that the rigidity was not constant as expected but was dependent on microtubule length. This length dependency explains the discrepancies among reported values of microtubule flexural rigidity measured by different methods. Comparing microtubules of identical lengths, microtubules assembled with brain-derived associated proteins (4 x 10(-23) Nm2 at around 10 microns in length) were four times more rigid than those assembled from purified tubulin and stabilized with taxol (1 x 10(-23) Nm2).

[1]  A. Matus,et al.  Actin depolymerisation induces process formation on MAP2-transfected non-neuronal cells. , 1993, Development.

[2]  R. Williams,et al.  Taxol-induced flexibility of microtubules and its reversal by MAP-2 and Tau. , 1993, The Journal of biological chemistry.

[3]  R. Himes,et al.  Alterations in number of protofilaments in microtubules assembled in vitro , 1978, The Journal of cell biology.

[4]  J. Mizushima-Sugano,et al.  Flexural rigidity of singlet microtubules estimated from statistical analysis of their contour lengths and end-to-end distances. , 1983, Biochimica et biophysica acta.

[5]  M. M. Bradford A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. , 1976, Analytical biochemistry.

[6]  J. Howard,et al.  Flexural rigidity of microtubules and actin filaments measured from thermal fluctuations in shape , 1993, The Journal of cell biology.

[7]  A. Ashkin,et al.  Optical trapping and manipulation of viruses and bacteria. , 1987, Science.

[8]  C. Cantor,et al.  Microtubule assembly in the absence of added nucleotides. , 1973, Proceedings of the National Academy of Sciences of the United States of America.

[9]  A C Maggs,et al.  Analysis of microtubule rigidity using hydrodynamic flow and thermal fluctuations. , 1994, The Journal of biological chemistry.

[10]  E. Nogales,et al.  Low resolution structure of microtubules in solution. Synchrotron X-ray scattering and electron microscopy of taxol-induced microtubules assembled from purified tubulin in comparison with glycerol and MAP-induced microtubules. , 1992, Journal of molecular biology.

[11]  R. Leighton,et al.  Feynman Lectures on Physics , 1971 .

[12]  A. Matus Stiff microtubules and neuronal morphology , 1994, Trends in Neurosciences.

[13]  R A Milligan,et al.  Kinesin follows the microtubule's protofilament axis , 1993, The Journal of cell biology.