MAP Kinase Signaling Protocols


Book Description

Mitogen-activated protein kinase (MAPK) signaling cascades are a group of protein kinases that play a central role in the intracellular transmission of extracellular signals. These cascades operate as major lines of communication within a complicated signaling network that regulates many cellular processes, including proliferation, differentiation, development, stress response, and apoptosis. More than 15,000 papers on MAPKs have been published over the past few years, with the number of publications increasing each year. More and more laboratories embark on the study of MAPK cascades in many d- tinct cellular systems and in particular their role in disease. Future challenges in the study of MAPK cascades remain in understa- ing the role of the various components and isoforms of the cascades in the multiple critical functions that they regulate in the whole organism, as well as the diseases caused by their malfunction. Data from gene-disrupted mice s- gest that inhibition of the MAPK cascades may have serious consequences on the development and growth of the animals. For example, targeted deletion of MEK1 is lethal, owing to developmental problems of placental vasculature and abnormal fibroblast migration. This lethality occurs in spite of the normal expression of MEK2, indicating that although the two MEK isoforms are apparently similar, they do have distinct functions, at least during embryog- esis. The ERK cascade was also shown to play a central role in brain function and in learning and memory.




MAP Kinase Signaling Protocols


Book Description

Mitogen-activated protein (MAP) kinase (MAPK) cascades are key signaling components that govern essentially all cellular processes evoked by any type of stimulation, and it has been well established that the malfunctioning of these cascades leads to various diseases including cancer, autoimmunity, and diabetes. In MAP Kinase Signaling Protocols, Second Edition, expert researchers fully update the popular first edition the key techniques used in the study of MAPK signaling cascades in various cellular contexts. This thorough volume explores essential topics such as activation and function of components of the MAPK signaling cascades, the study of MAPK cascades as transmitters of membranal receptor signals, structure-function relationships of MAPKs, studies on the regulation of MAPK cascades, the use of lower organisms, animal models, and human genetics in the study of MAPKs, as well as the study of MAPKs in specific systems and diseases. Written in the highly successful Methods in Molecular BiologyTM series format, chapters includes introductions to their respective subjects, lists of the necessary materials, step-by-step, readily reproducible laboratory protocols, and tips on troubleshooting and avoiding known pitfalls. Comprehensive and cutting-edge, MAP Kinase Signaling Protocols, Second Edition aims to facilitate the study of MAPKs and allow for quicker progress in our knowledge of many vital cellular processes as well as devastating diseases.




Stress-Activated Protein Kinases


Book Description

In this book leading researchers in the field discuss the state-of-the-art of many aspects of SAPK signaling in various systems from yeast to mammals. These include various chapters on regulatory mechanisms as well as the contribution of the SAPK signaling pathways to processes such as gene expression, metabolism, cell cycle regulation, immune responses and tumorigenesis. Written by international experts, the book will appeal to cell biologists and biochemists.




Transmembrane Signaling Protocols


Book Description

The previous edition of Transmembrane Signaling Protocols was published in 1998. Since then the human genome has been completely sequenced and new methods have been developed for the use of microarrays and proteomics to analyze global changes in gene expression and protein profiles. These advances have increased our ability to understand transmembrane signaling processes in much greater detail. They have also simultaneously enhanced our ability to determine the role of a large number of newly identified molecules in signaling events. In addition, novel video microscopy methods have been developed to image transmembrane signaling events in live cells in real time. In view of these major advances, it is time to update the previous edition. Because of the success of that volume, we have chosen to keep the essential character of the book intact. Introductory chapters from experts have been included to provide overall perspective and an overview of recent advances in signal transduction pathways. The individual chapters now include comp- hensive detailed methods, studies in genetically tractable systems, fluorescence microscopy in live single cells, ex vivo analysis of primary cells from tra- genic mice, as well as genomic and proteomic approaches to the analysis of transmembrane signaling events. We would like to express our deep gratitude to the coauthors of this publi- tion. We hope that Transmembrane Signaling Protocols, Second Edition will serve as a valuable resource for future progress in the study of signal transd- tion pathways.




Transmembrane Signaling Protocols


Book Description

This collection of practical, cutting-edge techniques for the study of cell signaling provides detailed, step-by-step instructions, helpful notes, and troubleshooting tips that make even the most powerful of the newest techniques readily reproducible. The protocols presented include the use of peptide libraries to study transmembrane signaling; the use of single-cell assays to analyze signal transduction pathways; the reconstitution of signaling complexes; methods for analyzing protein-protein interactions, and more. Introductory reviews explain the basic theory and enable researchers new to the area to rapidly gain understanding, as well as command of the practical knowledge and expertise afforded by the protocols. Transmembrane Signaling Protocols makes available to all researchers the many state-of-the-art techniques that have recently led to landmark discoveries in transmembrane signaling.




G Protein Signaling


Book Description

Alan V. Smrcka presents a collection of cutting-edge methods for investigating G protein signaling from a variety of perspectives ranging from in vitro biochemistry to whole animal studies. Among the readily reproducible techniques presented are those for the purification of G proteins and effectors enzymes, assays of these purified G proteins and effector enzymes, and for the study of G proteins interactions with effectors in intact cells. Additional methods are provided for assaying G protein coupled receptor structure, function, and localization, and for studying the physiological roles for endogenous G proteins.




Regulation of G1 Phase Progression


Book Description

In this contribution, several specialists describe the current knowledge on the molecular networks that regulate cell cycle progression, with an emphasis on the G1 phase of the cell cycle. The first part of Regulation of G1 Phase Progression is concerned with the individual molecules that form the network, including cyclins, cyclin-dependent kinases, inhibitors of these kinases and retinoblastoma and p53. The second section describes the signaling cascades by which external factors influence the cell cycle network, including mitogens, the extracellular matrix, nutrients and oxygen radicals. The last section describes the effects of specific external conditions on cell cycle progression and are presented such as serum starvation and subsequent re-addition and stress conditions (heat, osmolarity). The final two chapters describe the relation between cell cycle progression with cell differentiation and with apoptosis.




Plant Pattern Recognition Receptors


Book Description

"This volume covers protocols on techniques ranging from MAMP isolations from diverse microorganisms, PRR identifications from different plant species, MAMP-PRR binding, and a series of signaling responses and events revealed by various biochemical, cellular, genetic and bioinformatic tools. 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, Plant Pattern Recognition Receptors: Methods and Protocolsaims to ensure successful results in the further study of this vital field." -- OCLC.




Guide to Yeast Genetics and Molecular Biology


Book Description

Guide to Yeast Genetics and Molecular Biology presents, for the first time, a comprehensive compilation of the protocols and procedures that have made Saccharomyces cerevisiae such a facile system for all researchers in molecular and cell biology. Whether you are an established yeast biologist or a newcomer to the field, this volume contains all the up-to-date methods you will need to study "Your Favorite Gene" in yeast. Basic Methods in Yeast Genetics**Physical and genetic mapping**Making and recovering mutants**Cloning and Recombinant DNA Methods**High-efficiency transformation**Preparation of yeast artificial chromosome vectors**Basic Methods of Cell Biology**Immunomicroscopy**Protein targeting assays**Biochemistry of Gene Expression**Vectors for regulated expression**Isolation of labeled and unlabeled DNA, RNA, and protein




Cytokine Protocols


Book Description

A collection of biochemical, cellular, and molecular techniques for unraveling and quantifying the events occurring between the initial contact of a cytokine at the membrane receptor and the eventual activation of gene transcription. The techniques used include the generation of transfectants, the immunohistochemical detection of cytokines in tissue sections, and optimized staining for cytoplasmic detection. Highlights include RT-PCR of small amounts of mRNA, in situ hybridization, biosensor analysis, measurement of biological activities and standardization, immunohistochemical and single-cell detection, and receptor isolation, characterization, and crystallization. Enjoy a quick and smooth introduction to the key methods used in cytokine research Use readily reproducible techniques that ensure successful experimental results Employ antisense-RNA, RT-PCR of small amounts of mRNA, and in situ hybridization.