We use atomic force microscopy (AFM) to study the structure and mechanical properties of microtubules on the nanometer scale. AFM allows us to observe these protein polymers in buffer, under physiologically relevant conditions. In our experiments, microtubules are reconstituted in vitro, immobilized on 3-aminopropyltriethoxysilane (APTES)-coated glass or mica, and imaged using intermittent contact AFM. We hope to obtain high resolution images of the microtubules and compare this to data acquired using complementary techniques. In addition, the local mechanical properties of microtubules will be measured using force versus distance data obtained with AFM. We will explore the difference in rigidity between different types of microtubules. Towards this end, we are synthesizing a non-hydrolyzable analogue of GTP, guanylyl-(α,β)-methylene-diphosphonate (GMPCPP). GMPCPP allows for the polymerization of stable microtubules without the addition of other stabilizing agents which could affect the rigidity of microtubules. These studies will provide a mechanical profile of the microtubule lattice, contributing to a more complete understanding of dynamic instability and the properties of native microtubules.
Back to Biochemistry Poster Session II
Back to The 33rd Northeast Regional Meeting (July 14-17, 2005)