Gelb Protein Lipidation Protocols

Erscheinungsjahr 2008
ISBN: 978-1-59259-264-7
Verlag: Humana Press
Format: PDF
Kopierschutz: 1 - PDF Watermark

E-Book, Englisch, Band 116, 240 Seiten, eBook

Reihe: Methods in Molecular Biology

Zielgruppe

Professional/practitioner

Autoren/Hrsg.

Gelb, Michael H.

Inhaltsverzeichnis

In Vitro Analysis of GPI Biosynthesis in Mammalian Cells.- Selection of Mammalian Cell Mutants in GPI Biosynthesis.- Analysis of the Cell-Surface Distribution of GPI-Anchored Proteins.- Imaging Fluorescence Resonance Energy Transfer as Probe of Membrane Organization and Molecular Associations of GPI-Anchored Proteins.- 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 ?-, ?-, and ?-Subunits.- Analysis of Lipids in Caveolae.- Analysis of the Carbohydrate Components of Glycosylphosphatidylinositol Structures Using Fluorescent Labeling.- Analysis of the Lipid Moiety of GPI Anchor in the Yeast Saccharomyces cerevisiae.- Rapid Identification of Cysteine-Linked Isoprenyl Groups by Metabolic Labeling with [3H]Farnesol and [3H]Geranylgeraniol.- Incorporation of Radiolabeled Prenyl Alcohols and Their Analogs into Mammalian Cell Proteins.- Reconstitution of Yeast Farnesyltransferase from Individually Purified Subunits.- Probing the Role of H-Ras Lipidation for Signaling Functions in Xenopus laevis Oocytes.- Fluorescence Measurement of Lipid-Binding Affinity and Interbilayer Transfer of Bimane-Labeled Lipidated Peptides.- Determination of the Kinetics of Intervesicle Transfer and Transbilayer Diffusion of Bimane-Labeled Lipidated Peptides.- Preparation and Assay of Myristoyl-CoA:Protein N-Myristoyltransferase.- Metabolic Labeling of Protein-Derived Lipid Thioesters in Palmitoyl-Protein Thioesterase-Deficient Cells.- Fatty Acid Analysis of Protein-Derived Lipid Thioesters Isolated from Palmitoyl-Protein Thioesterase-Deficient Cells.