This new volume of Methods in Enzymology continues the legacy of this premier serial with quality chapters authored by leaders in the field. This volume covers cytoskeletal structure, including such topics as rotational movement of formins studied by fluorescence polarization microscopy, in vitro assembly assays for bacterial actin filaments, and modulators of microtubule plus end growth.
This new volume of Methods in Enzymology continues the legacy of this premier serial with quality chapters authored by leaders in the field. This volume covers cytoskeletal structure, including such topics as rotational movement of formins studied by fluorescence polarization microscopy, in vitro assembly assays for bacterial actin filaments, and modulators of microtubule plus end growth.
Polymer dynamics 1. Actin filament dynamics using microfluidics 2. Bacterial actin-like proteins: purification and characterization of self-assembly properties 3. Quantitative analysis of microtubule self-assembly kinetics and tip structure
Polymer nucleation and regulation 4. Biochemical reconstitution of the WAVE regulatory complex 5. Rotational movement of formins evaluated by using single-molecule fluorescence polarization 6. Single molecule studies of actin assembly and disassembly factors 7. Assaying microtubule nucleation by the ?-tubulin ring complex 8. Reconstituting dynamic microtubule polymerization regulation by TOG domain proteins. 9. Generation of differentially modified microtubules using in vitro enzymatic approaches
Molecular motor ensembles on natural and engineered cargoes 10. Engineering defined motor ensembles with DNA origami 11. Construction and analyses of elastically-coupled multiple motor systems 12. Reconstitution of cortical dynein function 13. Reconstitution of microtubule-dependent organelle transport 14. Reconstituting the motility of isolated intracellular cargoes
Building higher order networks and interactions 15. Reconstitution of contractile actomyosin arrays 16. Directed actin assembly and motility 17. In vitro reconstitution of dynamic microtubules interacting with actin filament networks 18. Measuring kinetochore-microtubule interaction in vitro 19. Micropattern-guided assembly of overlapping pairs of dynamic microtubules
Cell extract systems 20. WAVE regulatory complex activation 21. Dissecting principles governing actin assembly using yeast extracts 23. Glycogen-supplemented mitotic cytosol for analyzing Xenopus egg microtubule organization 24. Spindle movements on immobilized chromatin micropatterns
Polymer dynamics 1. Actin filament dynamics using microfluidics 2. Bacterial actin-like proteins: purification and characterization of self-assembly properties 3. Quantitative analysis of microtubule self-assembly kinetics and tip structure
Polymer nucleation and regulation 4. Biochemical reconstitution of the WAVE regulatory complex 5. Rotational movement of formins evaluated by using single-molecule fluorescence polarization 6. Single molecule studies of actin assembly and disassembly factors 7. Assaying microtubule nucleation by the ?-tubulin ring complex 8. Reconstituting dynamic microtubule polymerization regulation by TOG domain proteins. 9. Generation of differentially modified microtubules using in vitro enzymatic approaches
Molecular motor ensembles on natural and engineered cargoes 10. Engineering defined motor ensembles with DNA origami 11. Construction and analyses of elastically-coupled multiple motor systems 12. Reconstitution of cortical dynein function 13. Reconstitution of microtubule-dependent organelle transport 14. Reconstituting the motility of isolated intracellular cargoes
Building higher order networks and interactions 15. Reconstitution of contractile actomyosin arrays 16. Directed actin assembly and motility 17. In vitro reconstitution of dynamic microtubules interacting with actin filament networks 18. Measuring kinetochore-microtubule interaction in vitro 19. Micropattern-guided assembly of overlapping pairs of dynamic microtubules
Cell extract systems 20. WAVE regulatory complex activation 21. Dissecting principles governing actin assembly using yeast extracts 23. Glycogen-supplemented mitotic cytosol for analyzing Xenopus egg microtubule organization 24. Spindle movements on immobilized chromatin micropatterns
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