Protein Lipidation ProtocolsIn 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. |
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Obsah
In Vitro Analysis of GPI Biosynthesis in Mammalian Cells | 1 |
Selection of Mammalian Cell Mutants in GPI Biosynthesis | 13 |
Analysis of the CellSurface Distribution of GPIAnchored Proteins | 23 |
Imaging Fluorescence Resonance Energy Transfer as Probe | 37 |
Purification of CaveolaeDerived Membrane Microdomains Containing | 51 |
Analysis of Lipids in Caveolae | 61 |
Analysis of the Carbohydrate Components | 73 |
Analysis of the Lipid Moiety of GPI Anchor in the Yeast Saccharomyces | 91 |
Incorporation of Radiolabeled Prenyl Alcohols and Their Analogs into | 125 |
Reconstitution of Yeast Farnesyltransferase from Individually Purified | 145 |
Probing the Role of HRas Lipidation for Signaling Functions in Xeno | 161 |
Fluorescence Measurement of LipidBinding Affinity and Interbilayer | 177 |
Determination of the Kinetics of Intervesicle Transfer and Transbilayer | 187 |
Metabolic Labeling of ProteinDerived Lipid Thioesters in Palmitoyl | 213 |
Fatty Acid Analysis of ProteinDerived Lipid Thioesters Isolated from | 221 |
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