Protein Lipidation Protocols

Přední strana obálky
Michael H. Gelb
Springer Science & Business Media, 1999 - Počet stran: 240
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.
 

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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
Index
229

Rapid Identification of CysteineLinked Isoprenyl Groups By Metabolic
107

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