Dresden 2009 – scientific programme

Parts | Days | Selection | Search | Downloads | Help

BP: Fachverband Biologische Physik

BP 15: Motor Proteins

BP 15.2: Talk

Wednesday, March 25, 2009, 14:45–15:00, ZEU 260

The Motility of Monomeric and Dimeric Variants of Eg5 studied in the Presence of the Kinesin-5-specific Inhibitor Monastrol — Stefan Lakämper, •Christina Thiede, Stefanie Reiter, Kerstin v. Roden, and Christoph Schmidt — 3. Physikalisches Institut, Georg-August-Universität, 37077 Göttingen

The homo-tetrameric motor-protein Eg5 from Xenopus laevis drives relative sliding of anti-parallel microtubules, most likely by the processive action of its two sets of dimeric motor domains at each end. As recently shown by Kwok et al. (NCB 2006) and Kapitein et al. (JCB 2008), tetrameric motors move on a single microtubule in a fashion including diffusional and directional episodes, while motors moving between anti-parallel microtubules act in a highly directional and processive fashion. We have studied the processive behavior of a dimeric chimera (Eg5Kin) carrying the Eg5-motor and neck-linker and the Kinesin-1 neck and stalk. While Eg5Kin displays essentially the same motile properties as a truncated Eg5 (Eg5-513 his, Krysziak et al., JBC 2006, Valentine et al., NCB, 2006) its processivity is 40x increased to about 240 consecutive 8nm-steps on average, at a velocity of 95 nm/s. With increasing monastrol concentrations we find a dose-dependent and cooperative reduction in run length, but not in speed, indicating that two monastrol molecules are required to terminate a processive run. To further study the allosteric effect of monastrol on the motility of Eg5-motors, we generated monomeric and dimeric Eg5-constructs and compared their surface gliding-velocities in the presence of increasing concentrations monastrol.