The growing interest in recent years in the anchoring to membranes of proteins by post translational modification is documented by the large number of publications which ap peared in this field. In September 1987, scientists from 10 countries from all over the world met in the resort village of Les Diablerets, Switzerland, to discuss the most recent advances made in this field. The sessions were devoted to the anchoring of membrane proteins by cova lent attachment of fatty acids and of glycophospholipids. The workshop brought together many scientists working on vastly different proteins such as the variant surface glycoprotein of Trypanosomes and antigens of the mammalian cells. The subject of the workshop unified many scientists who had not met before and thus greatly stimulated interdisciplinary work. In addition to the lectures, each participant was provided with a collection of Methods currently in use in the study of membrane proteins anchored by post-translational modifica tion. An updated version of this collection is now presented as a Laboratory Manual, and the techniques described therein will give researchers easy and practical access to the investiga tion of post-translationally modified proteins. The publication of the present book by Springer follows an established tradition of previously published manuals on the handling of membrane proteins. Our thanks go to the authors who made the essential contribution in writing and adapting the experimental protocols, to Mrs. R.

Identification of Glycosyl-Phosphatidylinositol Membrane Anchors by Fatty Acid Labeling

[3H]Myristate-Labeled Variant Surface Glycoprotein from Trypanosoma brucei: Preparation and Use in the Assay of Glycan-Phosphatidylinositol-Specific Lipases
Identification of the Glycolipid Anchor of Alkaline Phosphatase by Metabolic Labeling
Reductive Radiomethylation and Amino Acid Analysis. A Sensitive Procedure to Identify Amine Components in Glycolipid Anchors of Membrane Proteins
Analytical and Preparative Phase Separation of Glycolipid-Anchored Membrane Proteins in Triton X-114 Solution
Assay and Purification of Pi-Specific Phospholipase C from Bacillus cereus Using Commercially Available Phospholipase C
Use of Phenylsepharose to Discriminate Between Hydrophilic and Hydrophobic Forms of Decay Accelerating Factor
PI-PLC Assay on Nitrocellulose Filter Immobilized Proteins
Deacylation and Reacylation of a Pure Enzyme to Demonstrate Covalent Glycerolipid
Incorporation of Rat Brain Thy-1 Antigen into Mouse T Lymphocytes
Identification and Characterization of Fatty Acid-Acylated Proteins in Cultured Cells by Radiolabeling
Chemical Analysis of Fatty Acids Covalently Attached to Proteins
Identification and Characterization of Proteins Undergoing Reversible Fatty Acid Acylation
Acylation of Proteins in Isolated Mitochondria
Plant Protein Acylation: Identification of the Modified Proteins and Analysis of the Bound Fatty Acid Ligand
Identification and Characterization of Lipid-Modified Membrane Proteins in Bacteria
Detection of Palmitoylating Activity with Exogenous Viral Acceptor Proteins
Lipid-Mediated Protein Glycosylation: Assembly of Lipid-Linked Oligosaccharides and Post-Translational Oligosaccharide Trimming
Immunochemical Characterization of the Ia Associated Invariant Chain
Acylation and Activation of Venom Phospholipase A2 Enzymes by Acyl Imidazolides
Isolation and Characterization of Bovine Erythrocyte Acetylcholinesterase Solubilized by PI-PLC
Characterization of Amphiphilic Forms of Cholinesterases by their Interactions with Non-Denaturing Detergents in Centrifugation and Charge-Shift Electrophoresis.