Protein Lipidation Protocols

1. Front Matter

   Pages i-x

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2. In Vitro Analysis of GPI Biosynthesis in Mammalian Cells

   • Victoria L. Stevens

   Pages 1-12

3. Selection of Mammalian Cell Mutants in GPI Biosynthesis

   • Victoria L. Stevens

   Pages 13-22

4. Analysis of the Cell-Surface Distribution of GPI-Anchored Proteins

   • Satyajit Mayor

   Pages 23-36

5. Imaging Fluorescence Resonance Energy Transfer as Probe of Membrane Organization and Molecular Associations of GPI-Anchored Proteins

   • Anne K. Kenworthy, Michael Edidin

   Pages 37-49

6. Purification of Caveolae-Derived Membrane Microdomains Containing Lipid-Anchored Signaling Molecules, Such as GPI-Anchored Proteins, H-Ras, Src-Family Tyrosine Kinases, eNOS, and G-Protein α-, β-, and γ-Subunits

   • Michael P. Lisanti, Massimo Sargiacomo, Philipp E. Scherer

   Pages 51-60

7. Analysis of Lipids in Caveolae

   • Jun Liu, Jan E. Schnitzer

   Pages 61-72

8. Analysis of the Carbohydrate Components of Glycosylphosphatidylinositol Structures Using Fluorescent Labeling

   • Nicole Zitzmann, Michael A. J. Ferguson

   Pages 73-89

9. Analysis of the Lipid Moiety of GPI Anchor in the Yeast Saccharomyces cerevisiae

   • Fulvio Reggiori, Elisabeth Canivenc-Gansel, Andreas Conzelmann

   Pages 91-105

10. Rapid Identification of Cysteine-Linked Isoprenyl Groups by Metabolic Labeling with [3H]Farnesol and [3H]Geranylgeraniol

    • Douglas A. Andres, Dean C. Crick, Brian S. Finlin, Charles J. Waechter

    Pages 107-123

11. Incorporation of Radiolabeled Prenyl Alcohols and Their Analogs into Mammalian Cell Proteins

    • Alberto Corsini, Christopher C. Farnsworth, Paul McGeady, Michael H. Gelb, John A. Glomset

    Pages 125-144

12. Reconstitution of Yeast Farnesyltransferase from Individually Purified Subunits

    • Jun Urano, Fuyuhiko Tamanoi

    Pages 145-159

13. Probing the Role of H-Ras Lipidation for Signaling Functions in Xenopus laevis Oocytes

    • Thomas Dudler, Michael H. Gelb

    Pages 161-176

14. Fluorescence Measurement of Lipid-Binding Affinity and Interbilayer Transfer of Bimane-Labeled Lipidated Peptides

    • John R. Silvius

    Pages 177-186

15. Determination of the Kinetics of Intervesicle Transfer and Transbilayer Diffusion of Bimane-Labeled Lipidated Peptides

    • John R. Silvius

    Pages 187-191

16. Preparation and Assay of Myristoyl-CoA:Protein N-Myristoyltransferase

    • Rajala V. S. Raju, Rajendra K. Sharma

    Pages 193-211

17. Metabolic Labeling of Protein-Derived Lipid Thioesters in Palmitoyl-Protein Thioesterase-Deficient Cells

    • Sandra L. Hofmann, Jui-Yun Lu

    Pages 213-219

18. Fatty Acid Analysis of Protein-Derived Lipid Thioesters Isolated from Palmitoyl-Protein Thioesterase-Deficient Cells

    • Sandra L. Hofmann, Linda A. Verkruyse

    Pages 221-228

19. Back Matter

    Pages 229-240

    PDF

In Protein Lipidation Protocols, Michael Gelb brings together a collection of readily reproducible techniques for studying protein lipidation, the covalent attachment of lipids to proteins. These cutting-edge methods-many never published before in a "hands-on" format-deal with glycosyl phosphatidylinositol (GPI)-containing compounds, protein fatty acylation, and protein prenylation. Included are novel techniques for determining the chemical structure of GPI-anchors, for radiolabeling the prenyl groups of protein in eukaryotic cells, a tool for developing inhibitors of the protein farnesyltransferase, and for an exciting lysosomal enzyme that cleaves fatty acyl groups from proteins, the first fatty acylase discovered.
Protein Lipidation Protocols offers biochemists, cell and molecular biologists, medicinal chemists, and pharmaceutical researchers state-of-the-art tools for understanding the complex biochemistry of protein lipidation, as well as catalyzing the development of many important new biopharmaceuticals, including anticancer drugs.

• University of Washington, Seattle

  Michael H. Gelb

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