Serpins: Methods and Protocols

Description

This volume explores the latest methods used to study and define serpin molecular structure, basic protease inhibition, serpin targets, and the roles of serpin in biology and disease using animal models. The chapters in this book cover topics such as crystallography and phage display, peptide design, phospholipid binding, and thrombus formation to microbiome analysis and development. 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 authoritative, Serpins: Methods and Protocols is a valuable resource for researchers and scientists interested in learning more about this evolving field. Front Matter ....Pages i-xiii Overview of Serpins and Their Roles in Biological Systems (Alexandra Lucas, Jordan R. Yaron, Liqiang Zhang, Sriram Ambadapadi)....Pages 1-7 Methods for Determining and Understanding Serpin Structure and Function: X-Ray Crystallography (Brian P. Mahon, Robert McKenna)....Pages 9-39 Serpin Phage Display: The Use of a T7 System to Probe Reactive Center Loop Libraries with Different Serine Proteinases (Lucas R. de Souza, Benjamin M. Scott, Varsha Bhakta, David A. Donkor, Darian L. Perruzza, William P. Sheffield)....Pages 41-64 Kinetic Measurement of Serpin Inhibitory Activity by Real-Time Fluorogenic Biochemical Assay (Jordan R. Yaron, Sriram Ambadapadi, Liqiang Zhang, Alexandra Lucas)....Pages 65-71 Methods for Identifying Virus-Derived Serpins (Masmudur M. Rahman)....Pages 73-86 In Vitro Approaches for the Assessment of Serpin Polymerization (Emma L. K. Elliston, David A. Lomas, James A. Irving)....Pages 87-107 Cellular Models for the Serpinopathies (Annamaria Fra, Emanuela D’Acunto, Mattia Laffranchi, Elena Miranda)....Pages 109-121 Binding of Serpins to Immobilized Phospholipids and Phospholipids in Suspension (Felix C. Wahlmüller, Judit Mihaly-Bison, Margarethe Geiger)....Pages 123-132 Viral Serpin Reactive Center Loop (RCL) Peptides: Design and Testing (Liqiang Zhang, Jordan R. Yaron, Sriram Ambadapadi, Alexandra Lucas)....Pages 133-142 In Vivo Analysis of Alpha-1-Antitrypsin Functions in Autoimmune Disease Models (Sihong Song, Yuanqing Lu, Ahmed S. Elshikha)....Pages 143-155 Analysis of In Vivo Serpin Functions in Models of Inflammatory Vascular Disease (Hao Chen, Sriram Ambadapadi, Erbin Dai, Liying Liu, Jordan R. Yaron, Liqiang Zhang et al.)....Pages 157-182 Gene Delivery of Alpha-1-Antitrypsin Using Recombinant Adeno-Associated Virus (rAAV) (Sihong Song, Yuanqing Lu)....Pages 183-196 Serpins in Venous Thrombosis and Venous Thrombus Resolution (Subhradip Mukhopadhyay, Tierra A. Johnson, Rajabrata Sarkar, Toni M. Antalis)....Pages 197-211 Next-Generation Sequencing Library Preparation for 16S rRNA Microbiome Analysis After Serpin Treatment (Juan Maldonado, Jordan R. Yaron, Liqiang Zhang, Alexandra Lucas)....Pages 213-221 Methods for Assessing Serpins as Neuroprotective Therapeutics (Jacek M. Kwiecien)....Pages 223-235 Adeno-Associated Virus Delivery of Viral Serpins for Ocular Diseases: Design and Validation (Cristhian J. Ildefonso, Alfred S. Lewin)....Pages 237-254 Serpins: Development for Therapeutic Applications (Alexandra Lucas, Jordan R. Yaron, Liqiang Zhang, Colin Macaulay, Grant McFadden)....Pages 255-265 Back Matter ....Pages 267-276