Characterization of Cell-Surface Material Released from AH-66 Hepatoma Ascites Cells into an Isotonic Solution

概要

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When AH-66 hepatoma ascites cells were incubated in an isotonic saline solution (PBS) at 0℃ for 1 h, material composed of approximately 90-95% protein and 5% carbohydrate was released without significantly affecting the viability of the cells. The material released into PBS contained no Na^+, K^+-adenosine triphosphatase and very low levels of phospholipid, cholesterol, and sialic acid, indicating the absence of contamination by fragments of the plasma membranes. The protein in the released material amounted to as much as approximately 2% of the total cell proteins and most of the protein components of the released material were identical with those of the plasma membranes as analyzed by sodium dodecyl sulfate-gel electrophoresis. The amino acid compositions of proteins of the released material were also very similar to those of the plasma membrane proteins. In contrast to the plasma membranes, however, the released material lacked periodate-Schiff-stainable glycoproteins and proteins with molecular weights larger than about 120000. The absence of sialic acid indicates that the carbohydrate chains, if any, of the proteins in the released material are incomplete. The surface structure of AH-66 cells implied by these results contrasts with that of erythrocyte membranes.
公益社団法人日本薬学会の論文
1986-01-25