Proteases and Cancer: Methods and Protocols

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. Front Matter ....Pages i-xv Dissecting Degradomes: Analysis of Protease-Coding Genes (Ángel Álvarez-Eguiluz, Ander Díaz-Navarro, Xose S. Puente)....Pages 1-13 Identification of Protease Cleavage Sites and Substrates in Cancer by Carboxy-TAILS (C-TAILS) (Nestor Solis, Christopher M. Overall)....Pages 15-28 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, Naohiko Koshikawa)....Pages 29-37 Biotin-Chasing Assay to Evaluate uPAR Stability and Cleavage on the Surface of Cells (Vladimir Leksa, Herbert B. Schiller, Hannes Stockinger)....Pages 39-47 Determination of Aconitase Activity: A Substrate of the Mitochondrial Lon Protease (Pedro M. Quirós)....Pages 49-56 A Simple Cell-Based Assay for the Detection of Surface Protein Shedding by Rhomboid Proteases (Angela Moncada-Pazos, Adam Graham Grieve)....Pages 57-64 Functional Production of Catalytic Domains of Human MMPs in Escherichia coli Periplasm (Dong Hyun Nam, Ki Baek Lee, Xin Ge)....Pages 65-72 Autophagy and Proteases: Basic Study of the Autophagic Flux by Western Blot (Álvaro F. Fernández)....Pages 73-81 Gel-Based Gelatin Zymography to Examine Matrix Metalloproteinase Activity in Cell Culture (Aastha Chhabra, Vibha Rani)....Pages 83-96 Analysis of Enzymatic Activity of Matrix Metalloproteinase (MMP) by Collagen Zymography in Melanoma (Vijay Walia, Yardena Samuels)....Pages 97-106 Measurement of Protease Activities Using Fluorogenic Substrates (Salvatore Santamaria, Hideaki Nagase)....Pages 107-122 Targeting the Expression of Cathepsin B Using CRISPR/Cas9 System in Mammalian Cancer Cells (Manu Gnanamony, Christopher S. Gondi)....Pages 123-131 Assessing the Influence of a Protease in Cell Migration Using the Barrier-Migration Assay (Tania Fontanil, Yamina Mohamedi, Santiago Cal, Álvaro J. Obaya)....Pages 133-143 Analysis of Invasive Activity of CAF Spheroids into Three Dimensional (3D) Collagen Matrices (María Ángeles Villaronga, Saúl Álvarez Teijeiro, Francisco Hermida-Prado, Marta Garzón-Arango, Victoria Sanz-Moreno, Juana María García-Pedrero)....Pages 145-154 Beyond the Proteolytic Activity: Examining the Functional Relevance of the Ancillary Domains Using Tri-Dimensional (3D) Spheroid Invasion Assay (Erico Tosoni Costa, Anamaria Aranha Camargo)....Pages 155-168 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 Klefström, Topi A. Tervonen)....Pages 169-178 Evaluation of Tumor Vasculature Using a Syngeneic Tumor Model in Wild-Type and Genetically Modified Mice (Francisco Javier Rodríguez-Baena, Silvia Redondo-García, María del Carmen Plaza-Calonge, Rubén Fernández-Rodríguez, Juan Carlos Rodríguez-Manzaneque)....Pages 179-192 3D Image Analysis of the Microvasculature in Healthy and Diseased Tissues (Álvaro Sahún-Español, Cristina Clemente, Alicia G. Arroyo)....Pages 193-212 Human Tumor Tissue-Based 3D In Vitro Invasion Assays (Pirjo Åström, Ritva Heljasvaara, Pia Nyberg, Ahmed Al-Samadi, Tuula Salo)....Pages 213-221 Ear Sponge Assay: A Method to Investigate Angiogenesis and Lymphangiogenesis in Mice (Maureen Van de Velde, Melissa García-Caballero, Tania Durré, Frédéric Kridelka, Agnès Noël)....Pages 223-233 Cancer Susceptibility Models in Protease-Deficient Mice (Alicia R. Folgueras, Sandra Freitas-Rodríguez, Yaiza Español, Gloria Velasco)....Pages 235-245 Imaging Proteolytic Activities in Mouse Models of Cancer (Anupama Pal, Alnawaz Rehemtulla)....Pages 247-260 Protease Silencing to Explore the Molecular Mechanisms of Cancer and Aging (Julia M. Fraile, Diana Campos-Iglesias, José M. P. Freije)....Pages 261-269 Analysis of Somatic DNA Methylation Alterations of Genes Encoding Cell Surface Metallopeptidases in Colorectal Cancer (Sergio Alonso, Beatriz González, Andreu Alibés, Manuel Perucho)....Pages 271-294 Targeting Deubiquitinases in Cancer (Joseph S. Bednash, Rama K. Mallampalli)....Pages 295-305 Generation of Highly Selective MMP Antibody Inhibitors (Dong Hyun Nam, Xin Ge)....Pages 307-324 Strategies to Target Matrix Metalloproteinases as Therapeutic Approach in Cancer (Zoi Piperigkou, Dimitra Manou, Konstantina Karamanou, Achilleas D. Theocharis)....Pages 325-348 Back Matter ....Pages 349-352