Title

Purification And Characterization Of An Endogenous Carbohydrate-Binding Protein From The Sea Urchin, Lytechinus Variegatus (eggs, Carrageenan, Development, Agar, Lectin)

Date of Award

1985

Availability

Article

Degree Name

Doctor of Philosophy (Ph.D.)

Department

Cell Biology and Anatomy

Abstract

A galactose-specific carbohydrate-binding protein has been purified to homogeneity from the eggs and embryos of the sea urchin, Lytechinus variegatus. In the unfertilized egg, this carbohydrate-binding protein comprises 0.1% of the total protein. The carbohydrate-binding protein displays microheterogeneity with an average isoelectric point of 7.1 by 2D SDS-PAGE. It is a glycoprotein, composed of 5.6% carbohydrate, including mannose, galactose, glucose, glucosamine, and galactosamine. This carbohydrate-binding protein binds to galactose as well as to polymers of galactose and galactose-sulfate (carrageenan) and polymers of fucose and fucose-sulfate (fucoidan). The carbohydrate-binding protein demonstrates a preference for A- over B-type bonds. Results from 1D SDS-PAGE suggest the carbohydrate-binding protein is a dimer of molecular weight 214,000D composed of equal sub-units of 118,000D. The carbohydrate-binding protein is predominantly cytoplasmic as demonstrated by its solubility properties. A specific antiserum was raised against the carbohydrate-binding protein and used in the immunofluorescent localization of the carbohydrate-binding protein. Prior to fertilization, the carbohydrate-binding protein was located in uniformly distributed aggregates throughout the cytoplasm. After fertilization, the aggregates containing this carbohydrate-binding protein migrate to the cell cortex. The carbohydrate-binding protein purified from L. variegatus was found to have some degree of immunologic cross reactivity with a lectin purified from the sea urchin, Anthocidaris crassispina suggesting similar proteins exist in other invertebrate eggs.

Keywords

Biology, General

Link to Full Text

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