The metabolism and functions of inositol phosphates impinge on various branches of biochemistry, physiology, and molecular biology, and methodological information is in consequence scattered far and wide. This book unites a selection of the most fundamental and commonly used techniques from leading international signal transduction laboratories, and brings together many valuable protocols for purifying and assaying inositides and related compounds. A novel feature is a catalogue of non-commercial sources of synthetic inositide analogues.
This text introduces the basic methodological tools to measure inositol lipids and phosphates and also describes new approaches that have become available in the last ten years, including g RNA-silencing and the protocols involving inositol pyrophosphates.
This new edition focuses on the applications of fluorescent and bioluminescent techniques for studying calcium signalling and calcium dependent processes. Cameleon probes, targeting bioluminescent probes, low affinity fluorescence indicators and chelators, and photometric techniques are all covered in depth. Also described in detail are methods for monitoring mitochondrial function, measuring calcium in the nuclear envelope and nucleoplasm, measuring calcium dependent gene expression, monitoring calcium signals in multicellular preparations, and measuring calcium extrusion. A major problem of fluorescent calcium measurements is the artifacts from zinc and transition metals and a chapter is devoted to their detection and minimization. Some of the pharmacological methods of studying calcium release are also covered. With many of the chapters written by the very scientists who developed the methods they are discussing this book is the most up-to-date, invaluable book for those studying cellular calcium.
This volume focuses on recent advances in the biochemical and molecular analysis of different families of phospholipases in plants and their roles in signaling plant growth, development and responses to abiotic and biotic cues. The hydrolysis of membrane lipids by phospholipases produces different classes of lipid mediators, including phosphatidic acid, diacylglycerol, lysophospholipids, free fatty acids and oxylipins. Phospholipases are grouped into different families and subfamilies according to their site of hydrolysis, substrate usage and sequence similarities. Activating one or more of these enzymes often constitutes an early, critical step in many regulatory processes, such as signal transduction, vesicular trafficking, secretion and cytoskeletal rearrangements. Lipid-based signaling plays pivotal roles in plant stress responses, cell size, shape, growth, apoptosis, proliferation, and reproduction.
This book provides descriptions of experimental methods in research on the cytoskeleton and its relationships to signaling and cell regulation. Thus, it bridges two active and fertile areas of research. The focus is directed particularly towards methods which take advantage of recent advances in molecular biology, microscopy and immunological assays. A second emphasis is on methods for understanding dynamic changes in cells. A third emphasis is on the formation and turnover of macromolecular and supramolecular complexes, which are so important in driving cell regulation and the behaviour of cytoskeletal elements. A combination of practical advice and detailed protocols should make this book valuable for both novice and experienced workers in these burgeoning fields.
The PI3Ks control many key functions in immune cells. PI3Ks phosphorylate PtdIns(4,5)P2 to yield PtdIns(3,4,5)P3. Initially, PI3K inhibitors such as Wortmannin, LY294002 and Rapamycin were used to establish a central role for Pi3K pathway in immune cells. Considerable progress in understanding the role of this pathway in cells of the immune system has been made in recent years, starting with analysis of various PI3K and Pten knockout mice and subsequently mTOR and Foxo knockout mice. Together, these experiments have revealed how PI3Ks control B cell and T cell development, T helper cell differentiation, regulatory T cell development and function, B cell and T cell trafficking, immunoglobulin class switching and much, much more. The PI3Kd inhibitor idelalisib has recently been approved for the treatment of B cell lymphoma. Clinical trials of other PI3K inhibitors in autoimmune and inflammatory diseases are also in progress. This is an opportune time to consider a Research Topic considering when what we have learned about the PI3K signalling module in lymphocyte biology and how this is making an impact on clinical immunology and haematology.
'Cell Signalling' presents a carefully structured introduction to this subject, introducing those conserved features which underlie many different extra-and intracellular signalling systems.
Since the publication of the first edition of Signal Transduction: A Practical Approach in 1992 there has been a great deal of new information about the processes of signal transduction and consequently many new methods have been developed. This new edition has therefore been updated and extended to include the major new methods now available. The first part of the book is mainly concerned with G protein-coupled receptors and covers structural studies of conformational changes and binding sites, phosphorylation and desensitisation, identification, receptor fusion proteins, and reporter gene systems. The second part includes methods for studying components of the other major families of signal transduction: adenylyl cylase and cAMP, phosphorylated inositol lipids, phosphinositide 3-kinases, phosphlipase D and phosphatidylcholine, sphingosine kinase, and inositol 1,4,5-triphosphate. Also included are chapters on baculoviral expression systems and the quantitative assay of mitogen activated protein kinases in intact cells and tissues. As with the previous edition Signal Transduction 2e covers a wide range of techniques and will be useful to both experienced researchers and newcomers.
This monograph explores the relationships between cell signaling and the cytoplasmic cytoskeleton in fundamental cell processes, thus bridging the gap between two very active aspects of molecular cell biology. It covers the two main - and reciprocal - questions of these relationships: How are structure and function of the cytoskeleton affected by external signals which impinge on the cell? How does the cytoskeleton influence the cellular signaling processes which determine cell behavior?
A comprehensive review of stress signaling in plants using genomics and functional genomic approaches Improving agricultural production and meeting the needs of a rapidly growing global population requires crop systems capable of overcoming environmental stresses. Understanding the role of different signaling components in plant stress regulation is vital to developing crops which can withstand abiotic and biotic stresses without loss of crop yield and productivity. Emphasizing genomics and functional genomic approaches, Protein Kinases and Stress Signaling in Plants is a comprehensive review of cutting-edge research on stress perception, signal transduction, and stress response generation. Detailed chapters cover a broad range of topics central to improving agricultural production developing crop systems capable of overcoming environmental stresses to meet the needs of a rapidly growing global population. This book describes the field of protein kinases and stress signaling with a special emphasis on functional genomics. It presents a highly valuable contribution in the field of stress perception, signal transduction and generation of responses against one or multiple stress signals. This timely resource: Summarizes the role of various kinases involved in stress management Enumerates the role of TOR, GSK3-like kinase, SnRK kinases in different physiological conditions Examines mitogen-activated protein kinases (MAPKs) in different stresses Describes the different aspects of calcium signaling under different stress conditions Examines photo-activated kinases (PAPKs) in varying light conditions Briefs the presence of tyrosine kinases in plants Highlights the cellular functions of receptor ]like protein kinases (RLKs) Possible implication of these kinases in developing stress tolerant crops Protein Kinases and Stress Signaling in Plants: Functional Genomic Perspective is an essential resource for researchers and students in the fields of plant molecular biology and signal transduction, plant responses to stress, plant cell signaling, plant protein kinases, plant biotechnology, transgenic plants and stress biology.
