图书简介
This volume provides a collection of techniques and approaches for the study of ERK signaling. It begins with a historical perspective of genetic and molecular discoveries, followed by chapters covering specific topics in a broad range of experimental systems, including in vitro assays of EGFR and ERK activities; proteomic and genome-wide analyses of ERK signaling targets; cell biological, genetic, quantitative and imaging approaches in cells and model organisms; and integrated mathematical modeling of the ERK pathway. Written in the highly successful Methods in Molecular Biology series format, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, and tips on troubleshooting and avoiding known pitfalls.
Cutting-edge and thorough, ERK Signaling: Methods and Protocols will aid and stimulate further advances in the vibrant field of ERK signaling.<
1. How Genetics has Helped Piece Together the MAPK Signaling Pathway Dariel Ashton-Beaucage and Marc Therrien 2. In Vitro Enzyme Kinetics Analysis of EGFR Zhihong Wang and Christine Candelora 3. High-Throughput Analysis of Mammalian Receptor Tyrosine Kinase Activation in Yeast Cells Nobuo Yoshimoto and Shun?ichi Kuroda 4. Structural Studies of ERK2 Protein Complexes Johannes F. Weijman, Stefan J. Riedl, and Peter D. Mace 5. Isolation and Characterization of Intrinsically Active (MEK-Independent) Mutants of Mpk1/Erk Tal Goshen-Lago, Dganit Melamed, Arie Admon, and David Engelberg 6. Assaying Activation and Subcellular Localization of ERK in Cells and Tissues Carme Caelles, Carles Bayod, and Melisa Morcillo 7. Detection and Functional Analysis of SUMO-Modified MEK Yuji Kubota and Mutsuhiro Takekawa 8. Single-Step Affinity Purification of ERK Signaling Complexes Using the Streptavidin-Binding Peptide (SBP) Tag Liu Yang and Alexey Veraksa 9. High-Throughput In Vitro Identification of Direct MAPK/Erk Substrates Rona Grossman and Ze?ev Paroush 10. Global Identification of ERK Substrates by Phosphoproteomics Based on IMAC and 2D-DIGE Hidetaka Kosako and Kou Motani 11. Analysis of Ras/ERK Compartmentalization by Subcellular Fractionation Lorena Agudo-Ibanez, Piero Crespo, and Berta Casar 12. Cell-Based Assays to Study ERK Pathway/Caveolin1 Interactions Raffaele Strippoli, Asier Echarri, Miguel Angel del Pozo 13. The Nuclear Translocation of ERK Denise A. Berti and Rony Seger 14. Visualization of RAS/MAPK Signaling In Situ by the Proximity Ligation Assay (PLA) Zijian Tang and Chengkai Dai 15. Measuring ERK Activity Dynamics in Single Living Cells Using FRET Biosensors Yannick Blum, Rafael D. Fritz, Hyunryul Ryu, and Olivier Pertz 16. Quantifying Tensile Force and ERK Phosphorylation on Actin Stress Fibers Hiroaki Hirata, Mukund Gupta, Sri Ram Krishna Vedula, Chwee Teck Lim, Benoit Ladoux, and Masahiro Sokabe 17. Co-Culture Activation of MAP Kinase in Drosophila S2 Cells Josefa Steinhauer 18. Isolation of Mouse Embryonic Stem Cell Lines in the Study of ERK1/2 MAP Kinase Signaling Marc K. Saba-El-Leil, Christophe Fremin, and Sylvain Meloche 19. 3D Organotypic Culture Model to Study Components of ERK Signaling Athina-Myrto Chioni, Rabia Tayba Bajwa, and Richard Grose 20. Genetic Validation of Cell Proliferation via Ras-Independent Activation of the Raf/Mek/Erk Pathway Carmen G. Lechuga, Lucia Simon-Carrasco, Harrys K.C. Jacob, and Matthias Drosten 21. Genome-Wide Analysis of RAS/ERK Signaling Targets Joshua P. Plotnik and Peter C. Hollenhorst 22. Probing Chromatin Modifications in Response to ERK Signaling Ozgur Oksuz and Wee-Wei Tee 23. Analyzing pERK Activation during Planarian Regeneration Susanna Fraguas, Yoshihiko Umesono, Kiyokazu Agata, and Francesc Cebria 24. Discovering Functional ERK Substrates Regulating Caenorhabditis elegans Germline Development Jessica Jie Chen and Swathi Arur 25. Reconstructing ERK Signaling in the Drosophila Embryo from Fixed Images Bomyi Lim, Carmeline J. Dsilva, Ioannis G. Kevrekidis, and Stanislav Y. Shvartsman 26.
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