Lipid Signaling Protocols / Edition 1

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Lipid Signaling Prools 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™ series format, the chapters provide readily reproducible laboratory prools, 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 Prools 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.

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Product Details

  • ISBN-13: 9781588297273
  • Publisher: Springer-Verlag New York, LLC
  • Publication date: 11/11/2008
  • Series: Methods in Molecular Biology Series, #462
  • Edition description: 2009
  • Edition number: 1
  • Pages: 429
  • Product dimensions: 6.30 (w) x 9.40 (h) x 1.00 (d)

Table of Contents

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-Ki Paik

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 Single and Two-Photon Fluorescence Lifetime Imaging Microscopy

Damien Alcor, Véronique Calleja and Banafshé Larijani

22) Phosphoinositide (PI) 3-kinase assays

Michael J. Fry

23) Measurement of phosphatidylinositol and phosphatidylcholine binding and transfer activity of the lipid transport protein PITP

Shamshad Cockcroft

24) In vitro reconstitution of activation of PLCe by Ras and Rho GTPases

Natalia Lamuño Gandarillas, Tom D. Bunney, Michelle B. Josephs, Peter Gierschik and Matilda Katan

25) Assaying endogenous Phosphatidylinositol-4-phosphate 5-kinase (PIP5K)


Jonathon R. Halstead, Mireille H. Snel, Sarah Meeuws, David R. Jones and Nullin Divecha

26) Preparation of Membrane Rafts

Mark G. Waugh and J. Justin Hsuan

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