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P9359 Sigma

Protein Lipidation Protocols

  •  ISBN-10 0-89603-534-4

  •  ISBN-13 978-0-89603-534-8

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publication info   M.H. Gelb, ed., Humana Press, 1999, 256 pp., hard cover
mfr. no.   Humana

Description

General description

This is a collection of reproducible techniques for studying protein lipidation. These methods, deal with glycosyl phosphatidylinositol (GPI)-containing compounds, protein fatty acylation, and protein prenylation. Included are 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 a lysosomal enzyme that cleaves fatty acyl groups from proteins.

Table of Contents

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 Linases, eNOS and G-Protein a-, b-, and g-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: A Useful Tool for Studying Protein Prenylation,
Reconstitution of Yeast Farnesyltransferase from Individually Purified Subunits.
Probing the Role of H-Ras Lipidation for Signaling Functions in Xenopus laevis Oocytes,
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,
Index.

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