Isolation and Chemical Characterization of a Highly Purified Phytomitogen from Phaseolus coccineus Seeds

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Abstract
An effective lymphocyte [rabbit] mitogen with a low erythroagglutinating ability isolated from the seeds of P. coccineus L. by ammonium sulfate precipitation and repeated ion-exchange chromatography on modified Sephadex. The mitogen stimulated lymphocytes at concentrations as low as 0.25 .mu.g/ml, the optimum concentration being in the range of 2.5-5.0 .mu.g/ml. This stimulating effect was completely inhibited by 5 mg/ml methyl .alpha.-D-mannopyranoside. Column gel filtration, disc electrophoresis on polyacrylamide gel in an alkaline medium and in the presence of sodium dodecylsulfate and the finding of a single N-terminal amino acid (alanine) showed the phytomitogen to be homogenous. Disc electrophoresis in an acid medium revealed the presence of 2 or 3 bands which are assumed to be due to aggregates of the mitogen. The minimum MW, as derived from sodium dodecylsulfate electrophoresis on polyacrylamide, was 35,000. The mitogen is a glycoprotein containing 10% neutral sugars and 1.3% amino sugars (glucosamine). The amino acid composition of the isolated phytomitogen is presented.

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