Glycoprotein Coat of the TA3 Cell. I. Removal of Carbohydrate and Protein Material From Viable Cells23

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Abstract
Incubation of viable TA3 ascites cells of the strain A mouse with neuraminidase released 0.44 mg of sialic acid per 109 cells, but no detectable protein in addition to that released in the absence of enzyme. This suggests that the reduced transplantability of the cells, after neuraminidase treatment, in the allogeneic C3H mouse is not due to the removal of nonspecific proteins from antigenic sites at the cell surface. Several methods for removing sialoglycopeptides, while maintaining cell viability, were studied. TPCK-trypsin or Pronase liberated carbohydrate and protein material weighing up to 15% of the dry weight of the cells and containing 71–74% of total cellular sialic acid. Ethylenediamine-tetraacetate released up to 11% of the cell weight, but less than 16% of the total sialic acid. Citrate was ineffective. All samples contained the same major carbohydrate components, mannose, galactose, glucose, N-acetylgalactosamine, Nacetylglucosamine, and sialic acid, but differed in the proportion of both carbohydrate and amino acid components. Successive trypsinizations of cells for short periods initially removed materials having high concentrations of sialic acid, hexosamine, and galactose. The sialic acid released by neuraminidase was composed of about 10% N-glycolylneuraminic acid and 90% N-acetylneuraminic acid.

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