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J. Biochem, 1980, Vol. 87, No. 4 1145-1151
© 1980 Japanese Biochemical Society

research-article

Isolation and Characterization of a Carbohydrate-Binding Protein from Dictyostelium discoideum NC-4 Deficient in Normal Discoidin Synthesis

Yukihiko ARAMAKI, Haruki YAMADA and Toshio MIYAZAKI

Department of Microbial Chemistry, Tokyo College of Pharmacy Hachioji, Tokyo 192-03

When Dictyostelium discoideum NC-4 cells are grown in conditions deficient for normal discoidin synthesis, cells synthesize a developmentally regulated carbohydrate-binding protein (CBP) which differs from discoidin in its properties. CBP was extracted with Tris-HCl buffer (pH 6.0), whereas it could not be extracted with the same buffer of pH 7.3 used for discoidin extraction. We compared the difference in the properties of CBP and discoidin. While formalinized rabbit erythrocytes were agglutinated by discoidin I and II, CBP did not agglutinate the same erythrocytes species. However, CBP had a high affinity to formalinized sheep erythrocytes. Agglutination of erythrocytes by CBP was strongly inhibited in the presence of N-acetyl-D-galactosamine, and specificity of the inhibitory sugar to agglutination was higher than discoidin. CBP was judged to be present on the cell surface, as Dictyostelium cells which were grown under the condition deficient for discoidin synthesis caused rosetts with sheep erythrocytes, and agglutination of erythrocytes with Dictyostelium cells was inhibited by the addition of N-acetyl-D-galactosamine.

Using formalinized sheep erythrocytes as an affinity absorbant, CBP was purified up to about 160-fold compared with crude extract in its specific activity. On polyacrylamide gel electrophoresis in the presence of sodium dodecyl sulfate, CBP gave a single protein band with a subunit molecular weight similar to one of the proteins in discoidin I. The pH stability of CBP was narrower than that of discoidin. This suggests that CBP may be important in the cell-cell adhesion of D. discoideum NC-4 cells, because the deficient cells of normal discoidin synthesis still retain the ability to form a normal aggregates.


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