Erk Signaling: Methods and Protocols

ISBN-13: 9781493964222 / Angielski / Twarda / 2016 / 440 str.

Gerardo Jimenez

Spis treści

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–Ibañez, 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 Frémin, 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, Lucía Simón-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 Cebrià

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. Drosophila

Marta Forés, Aikaterini Papagianni, Laura Rodríguez-Muñoz, and Gerardo Jiménez

27. Analyzing ERK Signal Dynamics during Zebrafish Somitogenesis

Takaaki Matsui and Yasumasa Bessho

28. Modeling RASopathies with Genetically Modified Mouse Models

Isabel Hernández-Porras and Carmen Guerra

29. Dissecting Cell-Fate Determination through Integrated Mathematical Modeling of the ERK/MAPK Signaling Pathway

Sung-Young Shin and Lan K. Nguyen

Blurb

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.

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