Adhesion Protein Protocols

The second edition of Adhesion Protein Protocols combines traditional techniques with cutting-edge and novel techniques that can be adapted easily to different molecules and cell types. The topics discussed include novel techniques for studying cell-cell adhesion, neutrophil chemotaxis, in vitro assays used to study leukocyte migration through monolayers of cultured endothelial cells, and novel techniques to purify pseudopodia from migratory cells. The protocols discussed in this volume are suitable for both novice and expert scientists, who will gain further insight into the complex and incompletely understood processes involved in cellular adhesion.

Integrins play pivotal roles not only across a wide range of physiological processes including tissue morphogenesis, immune responses, wound healing, and regulation of cell growth and differentiation, but also in numerous pathological phenomena such as autoimmunity, thrombosis, and cancer metastasis/progression. Therefore, investigations on integrins often demand multi-disciplinary approaches, making researchers long for a handy collection of comprehensive and practical protocols that detail experimental methods for studying integrin and related cell adhesion molecule functionality. Integrin and Cell Adhesion Molecules: Methods and Protocols aims to provide readers not only with basic protocols in studying integrin functions, but also with summaries on those state-of-the-art technologies that have been utilized for understanding integrin functionality at the cellular, molecular, structural, and organismal levels. Divided into six convenient sections, this detailed volume covers basic protocols for the study of integrin and related cell adhesion molecule functionality in vitro, illustrates structural biology approaches for studying integrins and related cell adhesion molecules, focuses on emerging imaging technologies for investigating cell migration, presents strategies to elucidate signaling through cell adhesion molecules, includes experimental techniques to investigate integrin functions at organismal levels in a physiological context, and showcases the most promising methods and technologies for the development of novel therapeutics and diagnostics. Written in the successful Methods in Molecular BiologyTM series format, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible protocols, and notes on troubleshooting and avoiding known pitfalls. Both experts and non-experts in the scientific community who wish to study cell adhesion molecules and diagnostics will find Integrin and Cell Adhesion Molecules: Methods and Protocols authoritative, easily accessible, and vastly informative.

Adhesion molecules are of fundamental importance in the regulation of immunity, inflammation, tissue remodeling, and embryonic development. They comprise different families of homologous proteins, such as selectins, integrins, cadherins, and immunoglobins. In addition, beyond these groups, other str- tures with adhesive properties, such as proteoglycans, occludin, and CD44, have been characterized recently. An understanding of the type and characteristics of adhesive molecules expressed by the different cell types and the possibility of manipulating their activity promises considerable clinical potential. Antibodies, small peptidic and nonpeptidic molecules, have recently been used to inhibit thrombosis by blocking platelet aggregation or inflammation through inhibition of leukocyte infiltration and adhesion. Inhibitors of adhesive molecules are used in expe- mental systems for the study of tumor growth and dissemination. Among major goals in the field are the identification of new members of the known adhesive protein families and of independent new adhesive structures. After structural characterization, even more demanding is the study of the biological activity of the new proteins, and the development of simple, rapid tests for the screening of possible inhibitors. In this regard, the production of such reagents as fragments and antibodies would help define the structure–function relati- ship of individual proteins. Data available in the literature show the complexity of the adhesive process and how different molecular epitopes might contribute to the adhesive properties of a single structure. Finally, a new area of investi- tion is the characterization of the intracellular signaling cascade triggered by the engagement of transmembrane adhesive proteins.

It is now widely accepted that much of the dynamic function of cells and tissues is regulated from outside the cell by the extracellular matrix. In ad- tion to its conventional role in providing a scaffold for building tissues, the extracellular matrix acts as a directional highway for cellular movement and provides instructional information for promoting survival, proliferation, and differentiation. Indeed, the extracellular matrix is beginning to take a starring role in the choreography of cell and tissue function. The diverse roles of the extracellular matrix are reflected in its highly complicated structure, consisting of an ever increasing number of components. Yet the mechanisms of extracellular matrix assembly and how they influences cell behavior are only just beginning to be understood. In order to solve these problems new methodologies are, of necessity, being developed. Many of these technologies are highly sophisticated and are currently available only in a ha- ful of laboratories. However, we believe that they can readily be transported and established by other researchers. Thus, the purpose of Extracellular Matrix Protocols is to present some of these complicated techniques in a style that is relatively easy to reproduce.

Cell adhesion plays a central role in development and disease. Cell adhesion to particular ligands can affect cytoskeletal organization and cell polarity, cell proliferation, and gene expression. This book is divided into two parts. The first section provides a discussion of the structure and function of the seven major classes of cell adhesion molecules: integrins, cadherins, selectins, heparan sulfate proteoglycans, the immunoglobulin superfamily, the ADAMs proteins, and transmembrane protein tyrosine phosphatases. The roles of these cell adhesion proteins in important processes such as cell mediated immunity, development and disease are discussed. In the second section, the molecular organization and function of junctional complexes, regions of the cell surface that are highly specialized for cell adhesion, are examined. Junctional complexes are now known to mediate adhesive interactions and contribute to transmembrane signaling events that dramatically influence cell behaviour. The biochemical organization of the adhesive membranes and the molecular mechanisms by which they transmit information to the cell are addressed. Written by contributors among the most prominent in the field, Cell Adhesion covers a wide range of topics in a single volume. It will be a great resource for students, teachers and researchers.

The second edition of Adhesion Protein Protocols combines traditional techniques with cutting-edge and novel techniques that can be adapted easily to different molecules and cell types. The topics discussed include novel techniques for studying cell-cell adhesion, neutrophil chemotaxis, in vitro assays used to study leukocyte migration through monolayers of cultured endothelial cells, and novel techniques to purify pseudopodia from migratory cells. The protocols discussed in this volume are suitable for both novice and expert scientists, who will gain further insight into the complex and incompletely understood processes involved in cellular adhesion.