Preface ......................................................... v
Contributors ................................................... xi
Part I Activation and Function of Components of the MAP
Kinase Signaling Cascades
1 The MAP Kinase Signaling Cascades: A System of Hundreds
of Components Regulates a Diverse Array of Physiological
Functions .................................................... 3
Yonat Keshet and Rony Seger
2 Determination of ERK Activity: Anti-phospho-ERK Antibodies
and In Vitro Phosphorylation ................................ 39
Shiri Procaccia, Sarah Kraus, and Rony Seger
3 Activation of SAPK/JNKs In Vitro ............................ 59
Deborah N. Chadee and John M. Kyriakis
4 Activation of p38 and Determination of Its Activity ......... 75
Huatnin Zhou, Jianming Chen, and Jiahuai Han
5 Activity Assays for Extracellular Signal-Regulated
Kinase 5 .................................................... 91
Kazuhiro Nakamura and Gary L. Johnson
6 Use of Inhibitors in the Study of MAP Kinases .............. 107
Kimberly Burkhard and Paul Shapiro
Part II Study of MAP Kinase Cascades as Transmitters of
Membranal Receptor Signals
7 MAP Kinase Activation by Receptor Tyrosine Kinases:
In Control of Cell Migration ............................... 125
Gabi Tarcic and Yosef Yarden
8 Activation of Ras and Rho GTPases and MAP Kinases
by G-Protein-Coupled Receptors ............................. 137
Mario Chiariello, Jose P. Vaqué, Piero Crespo, and
J. Silvio Gutkind
9 Regulation of MAP Kinase Signaling by Calcium .............. 151
Colin D. White and David B. Sacks
10 Identification of Novel Substrates of MAP Kinase Cascades
Using Bioengineered Kinases that Uniquely Utilize Analogs
of ATP to Phosphorylate Substrates ......................... 167
Hui Zheng, Adnan Al-Ayoubi, and Scott T. Eblen
11 ERK-MAP Kinase Signaling in the Cytoplasm .................. 185
Michelle C. Mendoza, Ekrem Emrah Er, and John Blenis
12 Lentiviral Vectors to Study the Differential Function of
ERK1 and ERK2 MAP Kinases .................................. 205
Marzia Indrigo, Alessand.ro Papale, Daniel Orellana, and
Riccardo Brambilla
Part III Structure-Function Relationships and Localization
of MAP Kinases
13 Structural Studies of MAP Kinase Cascade Components ........ 223
Elizabeth J. Goldsmith, Xiaoshan Min, Haixia He, and
Tianjun Zhou
14 Analysis of MAP Kinases by Hydrogen Exchange Mass
Spectrometry ............................................... 239
Kevin M. Sours and Natalie G. Ahn
15 A "Molecular Evolution" Approach for Isolation of
Intrinsically Active (MEK-Independent) MAP Kinases ......... 257
Vered Levin-Salomon, Oded Livnah, and David Engelberg
16 Reconstitution of the Nuclear Transport of the MAP Kinase
ERK2 ...................................................... 273
Arifjivan, Aarati Ranganatban, and Melanie H. Cobb
17 Localization and Trafficking of Fluorescently Tagged ERK1
and ERK2 ................................................... 287
Matilde Marchi, Riccardo Parra, Mario Costa, and Gian
Michele Ratto
Part IV Studies on the Regulation of MAP Kinase Cascades
18 Studying the Regulation of MAP Kinase by MAP Kinase
Phosphatases In Vitro and in Cell Systems .................. 305
Céline Tárrega, Caroline Nunes-Xavier, Rocío Cejudo-Marín,
Jorge Martin-Pérez, and Rafael Pulido
19 Proteomic Analysis of Scaffold Proteins in the ERK
Cascade .................................................... 323
Melissa M. McKay and Deborah K. Morrison
20 Analysis of ERKs' Dimerization by Electrophoresis .......... 335
Adán Pinto and Piero Crespo
21 MAP Kinase: SUMO Pathway Interactions ...................... 343
Shen-Hsi Yang and Andrew D. Sharrocks
22 Computational Modelling of Kinase Signalling Cascades ...... 369
David Gilbert, Monika Heiner, Rainer Breitling, and
Richard Orton
Part V Use of Lower Organisms, Animal Models, and Human
Genetics in the Study of MAP Kinases
23 Analysis of Mitogen-Activated Protein Kinase Activity in
Yeast ...................................................... 387
Elaine A. Elion and Rupam Sahoo
24 Detection of RTK Pathway Activation in Drosophila Using
Anti-dpERK Immunofluorescence Staining ..................... 401
Aharon Helman and Ze'ev Paroush
25 Studying MAP Kinase Pathways During Early Development of
Xenopus laevis ............................................. 409
Aviad Keren and Eyal Bengal
26 Deciphering Signaling Pathways In Vivo: The
Ras/Raf/Mek/Erk Cascade .................................... 421
Gergana Galabova-Kovacs and Manuela Baccarini
27 Mutational and Functional Analysis in Human Ras/MAP Kinase
Genetic Syndromes .......................................... 433
William E. Tidyman and Katherine A. Rauen
Part VI Study of MAP Kinases in Specific Physiological Systems
28 Implication of the ERK Pathway on the Post-transcriptional
Regulation of VEGF mRNA Stability .......................... 451
Khadija Essafi-Benkhadir, Jacques Pouysségur, and Gilles
Pagès
29 Studies on MAP Kinase Signaling in the Immune System ....... 471
Hongbo C.M. and Richard A. Flavell
30 Methods to Study MAP Kinase Signalling in the Central
Nervous System ............................................. 481
Bettina Wagner and Maria Sibilia
31 MAP Kinase Regulation of the Mitotic Spindle Checkpoint .... 497
Eva M. Eves and Marsha Rich Rosner
32 Using High-Content Microscopy to Study Gonadotropin
Releasing Hormone Regulation of ERK ........................ 507
Christopher J. Caunt, Stephen P. Armstrong, and Craig
A. McArdle
Index ......................................................... 525
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