Protein Tyrosine Phosphatases: Methods and Protocols

ISBN-13: 9781493937448 / Angielski / Twarda / 2016 / 402 str.

Rafael Pulido

Opis

This book provides coverage, methodology, and laboratory protocols on the more essential aspects of protein tyrosine phosphatase (PTP) function and regulation, including the use of standardized in vitro functional assays, suitable cell systems, and animal and microorganism models. Chapters covering state-of-the-art technical approaches suitable to decipher the physiologic roles of PTPs, and their involvement in tissue-specific functions, are also included, which will be of utility for both newcomers and experienced researchers in the field of tyrosine- and phosphoinositide- phosphorylation/dephosphorylation. 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. Authoritative and practical, Protein Tyrosine Phosphatases: Methods and Protocols aims to aid researchers in better defining the common and individual features of the PTP family members and translating this knowledge into PTP-based therapy for human disease.

Spis treści

1. The Extended Family of Protein Tyrosine Phosphatases

Andrés Alonso, Caroline E. Nunes-Xavier, Yolanda Bayón, and Rafael Pulido

2. Global RT-PCR and RT-qPCR Analysis of the mRNA Expression of the Human PTPome

Caroline E. Nunes-Xavier and Rafael Pulido

3. Expression, Purification, and Kinetic Analysis of PTP Domains

Mihaela Mentel, Rodica A. Badea, Georgiana Petrareanu, Sujay T. Mallikarjuna, Aura E. Ionescu, and Stefan E. Szedlacsek

4. Peptide Microarrays for Real Time Kinetic Profiling of Tyrosine Phosphatase Activity of Recombinant Phosphatases and Phosphatases in Lysates of Cells or Tissue Samples

Liesbeth Hovestad-Bijl, Jeroen van Ameijde, Dirk Pijnenburg, Riet Hilhorst, Rob Liskamp, and Rob Ruijtenbeek

5. Tailor-Made Protein Tyrosine Phosphatases: In Vitro Site-Directed Mutagenesis of PTEN and PTPRZ-B

Sandra Luna, Janire Mingo, Olaia Aurtenetxe, Lorena Blanco, Laura Amo, Jan Schepens, Wiljan J. Hendriks, and Rafael Pulido

6. Assays to Measure PTEN Lipid Phosphatase Activity In Vitro from Purified Enzyme or Immunoprecipitates

Laura Spinelli and Nicholas R. Leslie

7. Assessing the Biological Activity of the Glucan Phosphatase Laforin

Carlos Romá-Mateo, Madushi Raththagala, Mathew S. Gentry, and Pascual Sanz

8. Discovery and Evaluation of PRL Trimer Disruptors for Novel Anticancer Agents

Yunpeng Bai, Zhi-Hong Yu, and Zhong-Yin Zhang

9. Analyzing Pseudophosphatase Function

Shantá D. Hinton

10. Crystallization of PTP Domains

Colin Levy, James Adams, and Lydia Tabernero

11. NMR Spectroscopy to Study MAP Kinase Binding to MAP Kinase Phosphatases

Wolfgang Peti and Rebecca Page

12. Visualizing and Quantitating the Spatiotemporal Regulation of Ras/ERK Signaling by Dual-Specificity Mitogen-Activated Protein Phosphatases (MKPs)

Christopher J. Caunt, Andrew M. Kidger, and Stephen M. Keyse

13. In Situ Proximity Ligation Assay (In Situ PLA) to Assess PTP-Protein Interactions

Sina Koch, Irene Helbing, Sylvia-Annette Böhmer, Makoto Hayashi, Lena Claesson-Welsh, Ola Söderberg, and Frank-D. Böhmer

14. Use of Dominant Negative/Substrate Trapping PTP Mutations to Search for PTP Interactors/Substrates

Vegesna Radha

15. Detection and Identification of Ligands for Mammalian RPTP Extracellular Domains

Andrew William Stoker

16. Production of Osteoclasts for Studying Protein Tyrosine Phosphatase Signaling

Eynat Finkelshtein, Einat Levy-Apter, and Ari Elson

17. Functional Analysis of Protein Tyrosine Phosphatases in Thrombosis and Haemostasis

Souad Rahmouni, Alexandre Hego, Céline Delierneux, Odile Wéra, Lucia Musumeci, Lutz Tautz, and Cécile Oury

18. Functional Analysis of Dual-Specificity Protein Phosphatases in Angiogenesis

Mathieu Amand, Charlotte Erpicum, Christine Gilles, Agnès Noël, and Souad Rahmouni

19. Studying Protein-Tyrosine Phosphatases in Zebrafish

Alexander James Hale and Jeroen den Hertog

20. Live staining of Drosophila Embryos with RPTP Fusion Proteins to Detect and Characterize Expression of Cell-Surface RPTP Ligands

Namrata Bali, Hyung-Kook (Peter) Lee, and Kai Zinn

21. Methods to Study Protein Tyrosine Phosphatases Acting on Yeast MAPKs

Almudena Sacristán-Reviriego, María Molina, and Humberto Martín