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.
- Continues the legacy of this premier serial with quality chapters authored by leaders in the field
- Covers cytoskeletal structure
- Contains chapters with such topics as reconstitution of organelle transport and assaying microtubule nucleation by gamma-tubulin ring complex.
Table of Contents
Section I - 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
Section II - 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 y-tubulin ring complex
8. Reconstituting dynamic microtubule polymerization regulation by TOG domain proteins.
9. Generation of differentially modified microtubules using in vitro enzymatic approaches
Section III - 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
Section IV - 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
Section V - Cell extract systems
20. WAVE regulatory complex activation
21. Dissecting principles governing actin assembly using yeast extracts
22. Xenopus egg cytoplasm with intact actin
23. Glycogen-supplemented mitotic cytosol for analyzing Xenopus egg microtubule organization
24. Spindle movements on immobilized chromatin micropatterns
Ronald D. Vale is Professor of the Dept. of Cellular and
Molecular Pharmacology at the University of California, San Francisco and is
an Investigator in the Howard Hughes Medical Institute.
Vale received B.A. degrees in Biology and Chemistry from the University of California, Santa Barbara in 1980. Dr. Vale received his Ph.D. in Neuroscience from Stanford University in 1985 where he trained with Dr. Eric Shooter. He was a Staff Fellow with the N.I.H. stationed at the Marine Biological Laboratory in 1985-6 (with Tom Reese) and began his faculty appointment at UCSF in 1987. During his time at UCSF, he has served as the Director of the Cell Biology Program (4 years) and Vice-Chair (10 years) and Chair of the Dept. of Cellular and Molecular Pharmacology (5 years). He also holds an Adjunct Senior Scientist appointment with the Marine Biological Laboratory at Woods Hole.
Vale has served the scientific community by his participation on an NIH study section (1996-2002; chairing from 2000-2002). He served as President of the American Society of Cell Biology and on their International Affairs Committee. Vale is active in many educational activities. He founded and produces iBioSeminars, lectures by leading biologists which are made freely available on the web. Vale co-directed the 7 week Physiology Course at the Marine Biological Laboratory, transforming it to a venue of interdisciplinary research in physics and biology (2004-2008). His laboratory developed MicroManager, a widely used open source and freely available software package for microscopy. He also developed an educational web site on microscopy for elementary school children. Vale is active in helping young scientists in India by starting the very popular Young Investigator Meetings as well as an interactive web site (IndiaBioscience.org) for India biologists to obtain information on jobs/grants/collaborations.
Dr. Vale’s awards include the Pfizer Award in Enzyme Chemistry (1991), the Young Investigator Award from the Biophysical Society (1993), the Wiley Prize in Biomedical Sciences (2012), and the Albert Lasker Award for Basic Medical Research (2012). He was elected to the National Academy of Sciences in 2001 and to the American Academy of Sciences in 2002.