PART 1. Lipid systems and techniques Introduction - by Mike Wakelam 1) Eicosanoids: generation and detection in mammalian cells Valerie B. O'Donnell, Ben Maskrey and Graham W. Taylor. 2) Lipidomic analysis of phospholipids and related structures by liquid chromatography-mass spectrometry Trevor R. Pettitt 3) Measurement of Polyphosphoinositides in Cultured Mammalian Cells Frank T. Cooke 4) Inositol lipid-dependent functions in Saccharomyces cerevisiae: analysis of phosphatidylinositol phosphates Stephen K. Dove and Robert H. Michell 5) Methods for the determination of the mass of nuclear PtdIns4P, PtdIns5P and PtdIns(4,5)P2 David R. Jones, Yvette Bultsma and Nullin Divecha 6) Lipid Quantification and Structure Determination of Nuclear Envelope Precursor Membranes in the Sea Urchin Marie Garnier-Lhomme, Erick J. Dufourc, Banafshé Larijani and Dominic Poccia 7) Solution and solid state NMR of lipids Axelle Grelard, Anthony Couvreux, Cécile Loudet and Erick J. Dufourc 8) Spatial localisation of PIP2 in phase separated giant unilamellar vesicles with a fluorescent PLC-delta 1 PH domain Xavier Mulet, Erika Rosivatz, Ka Kei Ho, Béatrice L.L.E. Gauthé, Oscar Ces, Richard H. Templer, Rudiger Woscholski 9) Studying cell-to-cell interactions: an easy method of tethering ligands on artificial membranes Sebastian J. Fleire and Facundo D. Batista 10) Ceramide-induced transbilayer (flip-flop) lipid movement in membranes F.-Xabier Contreras, Ana-Victoria Villar, Alicia Alonso and Félix M. Goñi 11) Effects of Sterols on the Development and Ageing of Caenorhabditis elegans Eun-Young Lee, Pan-Young Jeong, Sun-Young Kim, Yhong-Hee Shim, and Young-KiPaik 12) Proteomic Analysis of the Sterol-Mediated Signalling Pathway in Caenorhabditis elegans Byung-Kwon Choi, Yun-Kyung Shin, Eun-Young Lee, Pan-Young Jeong, Yhong-Hee Shim and Young-Ki Paik PART 2. Lipid metabolism and recognition Introduction - by Robin Irvine 13) Studying the subcellular localisation and DNA-binding activity of FoxO transcription factors, downstream effectors of PI3K/Akt. Abdelkader Essafi, Ana R. Gomes, Karen M. Pomeranz, Aleksandra K. Zwolinska, Rana Varshochi, Ursula B. McGovern and Eric W.-F. Lam 14) Measurement of PTEN Activity In Vivo by Imaging Phosphorylated Akt Erika Rosivatz and Rudiger Woscholski 15) Analysis of phosphatidylinositol 3,4,5 trisphosphate 5-phosphatase activity by in vitro and in vivo assays Lisa M. Ooms, Jennifer M. Dyson, Anne M. Kong and Christina A. Mitchell 16) Measuring Phospholipase D activity in Insulin Secreting Pancreatic ß-cells and Insulin Responsive Muscle Cells and Adipocytes Rosanna Cazzolli, Ping Huang, Shuzhi Teng and William E. Hughes 17) Protein Kinase C as an effector of lipid derived second messengers Marie-Hélène Paclet, Jan K. Davidson-Moncada, Guillermo López-Lluch, Dongmin Shao and Lodewijk V Dekker 18) Detection of myotubularin phosphatases activity on phosphoinositides in vitro and ex vivo Holger Maria Rohde, Hélène Tronchère, Bernard Payrastre and Jocelyn Laporte 19) Quantification of Multiple Phosphatidylinositol 4-Kinase Isozyme Activities in Cell Extracts Mark G. Waugh, Shane Minogue and J. Justin Hsuan 20) Phospholipid-interacting proteins by solution state NMR spectroscopy Keiichiro Kami, Sundaresan Rajesh and Michael Overduin 21) Revealing Signalling in Single Cells by
Lipid Signaling Protocols assembles in a single volume the various tools and methodologies needed by the interested investigator to unravel lipid dependent signaling and cell function. Divided into two convenient sections, the volume begins by summarizing the physical properties of hydrophobic metabolites as well as the physical methodologies used for their analysis, which leads to the second section and its selection of biological methods, focused around the most relevant lipids, their corresponding metabolizing enzymes and the recognition proteins. Following the highly successful Methods in Molecular Biology(TM) series format, the chapters provide readily reproducible laboratory protocols, lists of necessary materials and reagents, and the tips on troubleshooting and avoiding known pitfalls.
Contributed to by top researchers in the field, Lipid Signaling Protocols is an essential resource for both experienced and novice researchers who desire a better understanding of the application of physical methodologies in the context of lipid signaling and lipid metabolism in cell biology.
Describes cutting-edge lipid analysis and quantification methods
Provides a selection of practical approaches to the study of lipid metabolism and recognition in the cellular context
Includes reproducible, step-by-step laboratory protocols
Chapters written by carefully selected experts in the field