A Membrane Glycoprotein of Aggregating Dictyostelium Cells with the Properties of Contact Sites
- 1 September 1979
- journal article
- Published by Wiley in European Journal of Biochemistry
- Vol. 99 (2), 419-426
- https://doi.org/10.1111/j.1432-1033.1979.tb13271.x
Abstract
Aggregating cells of Dictyostelium discoideum form EDTA-stable contacts which are blocked by a Fab (antigen-binding fragment) preparation from antisera raised against membranes. The target site of the blocking Fab fragments has been identified as a specific glycoprotein. In this paper its purification, carbohydrate and amino acid composition are described. Purification was 800-fold, starting with cells lysed by digitonin. The plasma membranes, preserved as ghosts by this treatment, were purified in a two-phase system and extracted with butan-1-ol. The water phase contained predominantly concanavalin-A-binding glycoproteins and was particularly rich in contact sites A. These were further purified on DE-cellulose and sucrose gradients. Sodium dodecylsulphate/polyacrylamide gel electrophoresis of the purified material revealed one major glycoprotein band in the molecular weight region of 80 000 to 90 000, depending on the acrylamide concentration. The sugars found in contact sites A were mannose, N-acetylglucosamine, fucose, and possibly glucose. The protein moeity contained 8% proline and was particularly rich in hydroxy amino acids.Keywords
This publication has 38 references indexed in Scilit:
- A specific glycoprotein as the target site of adhesion blocking Fab in aggregating Dictyostelium cellsNature, 1978
- Implication of developmentally regulated Concanavalin A binding proteins of Dictyostelium in cell adhesion and cyclic AMP regulationCell Differentiation, 1977
- The effects of proteases on proteins and glycoproteins of Dictyostelium discoideum plasma membranesExperimental Cell Research, 1977
- Cell surface location of discoidin, a developmentally regulated carbohydrate-binding protein from Dictyostelium discoideumExperimental Cell Research, 1975
- Solubilized contact sites a from cell membranes of Dictyostelium discoideumFEBS Letters, 1975
- Glycosphingolipids and Glycoproteins in the Wild‐Type and in a Non‐Aggregating Mutant of Dictyostelium discoideumEuropean Journal of Biochemistry, 1974
- Defective cyclic adenosine-3′,5′-phosphate-phosphodiesterase regulation in morphogenetic mutants of Dictyostelium discoideumJournal of Molecular Biology, 1973
- A rapid method for the isolation of L-cell surface membranes using an aqueous two-phase polymer systemThe Journal of Membrane Biology, 1971
- Cleavage of Structural Proteins during the Assembly of the Head of Bacteriophage T4Nature, 1970
- Immunochemical Studies on Core Lipopolysaccharides of Enterobacteriaceae of Different GeneraEuropean Journal of Biochemistry, 1970