J active substance associated with protein was purified from cattle Jcs serum extracted with lipid solvents. Serum residue, after extraction, was deproteinized first by heat coagulation and then by extraction with Genetron. Fractionation of the resulting deproteinized J active material was performed on DEAE-Sephadex, and the most active fraction eluted from this column with 0.15 M NaCl, was finally purified by preparative polycrylamide gel electrophoresis. Purified J active material exhibited a single band on analytical polyacrylamide gel, and was composed of protein, carbohydrate, long-chain base, phosphorus, glycerol and fatty acid. This purified protein-associated J substance is eight fold less active than the previously described J lipid when tested by hemolysis-inhibition assay with the J system. The decrease in activity parallels the decrease in phosphosphingoglycolipid content in the protein-associated J substance. The capacity to inhibit hemolysis (J system) was lost when the substance was subjected to mild alkaline methanolysis, periodate-oxidation or action by phospholipase C. Enzymatic desialytion did not affect the activity of the protein-bound J substance. Neither the J active lipid nor the protein-associated substance inhibited agglutination in the human A system.
All Science Journal Classification (ASJC) codes