Main description:

This volume explores the role of proteases in various catalytic classes and cellular processes associated with cancer. The chapters in this book cover topics such as encoding proteolytic enzymes, cleavage sites of proteases, functional catalytic domains of MMPs, zymography, angiogenesis and lymphangiogenesis, genetic and epigenetic alterations of metallopeptidases, and methods to target metalloproteinases. 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.

Comprehensive and cutting edge, Proteases and Cancer: Methods and Protocols is a valuable resource for clinical researchers across numerous fields such as molecular and cellular biology, oncology, biochemistry, and chemistry. This book is also useful for PhD students in the various disciples of health sciences.

Contents:

Preface...
Table of Contents...
Contributing Authors...

1. Dissecting Degradomes: Analysis of Protease-Coding Genes
Angel Alvarez-Eguiluz, Ander Diaz-Navarro, and Xose S. Puente

2. Identification of Protease Cleavage Sites and Substrates in Cancer by Carboxy-TAILS (C-TAILS)
Nestor Solis and Christopher M. Overall

3. Identification of Proteolytic Cleavage Sites of EphA2 by Membrane Type-1 Matrix Metalloproteinase on the Surface of Cancer Cells
Keiji Kikuchi, Hiroko Kozuka-Hata, Masaaki Oyama, Motoharu Seiki, and Naohiko Koshikawa

4. Biotin Chasing Assay to Evaluate uPAR Stability and Cleavage on the Surface of Cells
Vladimir Leksa, Herbert B. Schiller, and Hannes Stockinger

5. Determination of Aconitase Activity: A Substrate of the Mitochondrial Lon Protease
Pedro M. Quiros

6. A Simple Cell-Based Assay for the Detection of Surface Protein Shedding by Rhomboid Proteases
Angela Moncada-Pazos and Adam G. Grieve

7. Functional Production of Catalytic Domains of Human MMPs in Escherichia coli Periplasm
Dong Hyun Nam, Ki Baek Lee, and Xin Ge

8. Autophagy and Proteases: Basic Study of the Autophagic Flux by Western Blot
Alvaro F. Fernandez

9. Gel-Based Gelatin Zymography to Examine Matrix Metalloproteinase Activity in Cell Culture
Aastha Chhabra and Vibha Rani

10. Analysis of Enzymatic Activity of Matrix Metalloproteinase (MMP) by Collagen Zymography in Melanoma
Vijay Walia and Yardena Samuels

11. Measurement of Protease Activities using Fluorogenic Substrates
Salvatore Santamaria and Hideaki Nagase

12. Targeting the Expression of Cathepsin B using CRISPR/Cas9 System in Mammalian Cancer Cells
Manu Gnanamony and Christopher S. Gondi

13. Assessing the Influence of a Protease in Cell Migration using the Barrier-Migration Assay
Tania Fontanil, Yamina Mohamedi, Santiago Cal, and Alvaro J. Obaya

14. Analysis of Invasive Activity of CAF-Spheroids into Three-Dimensional (3D) Collagen Matrices
Maria Angeles Villaronga, Saul Alvarez Teijeiro, Francisco Hermida-Prado, Marta Garzon-Arango, Victoria Sanz-Moreno, and Juana Maria Garcia-Pedrero

15. Beyond the Proteolytic Activity: Examining the Functional Relevance of the Ancillary Domains using Tri-Dimensional (3D) Spheroid Invasion Assay
Erico Tosoni Costa and Anamaria Aranha Camargo

16. Analyzing the Type II Transmembrane Serine Protease Hepsin-Dependent Basement Membrane Remodeling in 3D Cell Culture
Shishir M. Pant, Denis Belitskin, Hanna Ala-Hongisto, Juha Klefstroem, and Topi A. Tervonen

17. Evaluation of Tumor Vasculature using a Syngeneic Tumor Model in Wild Type and Genetically Modified Mice
Francisco Javier Rodriguez-Baena, Silvia Redondo-Garcia, Maria del Carmen Plaza-Calonge, Ruben Fernandez-Rodriguez, and Juan Carlos Rodriguez-Manzaneque

18. 3D Image Analysis of the Microvasculature in Healthy and Diseased Tissues
Alvaro Sahun-Espanol, Cristina Clemente, and Alicia G. Arroyo

19. Human Tumor Tissue-Based 3D In Vitro Invasion Assays
Pirjo Astroem, Ritva Heljasvaara, Pia Nyberg, Ahmed Al-Samadi, and Tuula Salo

20. Ear Sponge Assay: A Method to Investigate Angiogenesis and Lymphangiogenesis in Mice
Maureen Van de Velde, Melissa Garcia-Caballero, Tania Durre, Frederic Kridelka, and Agnes Noel

21. Cancer Susceptibility Models in Protease-Deficient Mice
Alicia R. Folgueras, Sandra Freitas-Rodriguez, Yaiza Espanol, and Gloria Velasco

22. Imaging Proteolytic Activities in Mouse Models of Cancer
Anupama Pal and Alnawaz Rehemtulla

23. Protease Silencing to Explore the Molecular Mechanisms of Cancer and Aging
Julia M. Fraile, Diana Campos-Iglesias, and Jose M. P. Freije

24. Analysis of Somatic DNA Methylation Alterations of Genes Encoding Cell Surface Metallopeptidases in Colorectal Cancer
Sergio Alonso, Beatriz Gonzalez, Andreu Alibes, and Manuel Perucho

25. Targeting Deubiquitinases in Cancer
Joseph S. Bednash and Rama K. Mallampalli

26. Generation of Highly Selective MMP Antibody Inhibitors
Dong Hyun Nam and Xin Ge

27. Strategies to Target Matrix Metalloproteinases as Therapeutic Approach in Cancer
Zoi Piperigkou, Dimitra Manou, Konstantina Karamanou, and Achilleas D. Theocharis