Chromophore-assisted light inactivation and self-organization of microtubules and motors

摘要: Chromophore-assisted light inactivation (CALI) offers the only method capable of modulating specific protein activities in localized regions and at particular times. Here, we generalize CALI so that it can be applied to a wider range tasks. Specifically, show work with genetically inserted epitope tag; investigate effectiveness alternative dyes, especially fluorescein, comparing them standard dye, malachite green; study relative efficiencies pulsed continuous-wave illumination. We then use fluorescein-labeled hemagglutinin antibody fragments, together relatively low-power illumination examine targeted kinesin. destroy kinesin activity least two ways: either result apparent loss motor activity, or cause irreversible attachment enzyme its microtubule substrate. Finally, apply this implementation an vitro system proteins microtubules is self-organized aster formation. In system, effectively perturb local structure formation by blocking reducing degree chosen sample, without influencing elsewhere.