Open AccessPURIFICATION OF MITOGENIC PROTEINS DERIVED FROM Phaseolus vulgaris : ISOLATION OF POTENT AND WEAK PHYTOHEMAGGLUTININS POSSESSING MITOGENIC ACTIVITY
Author(s) -
Lawrence W. Allen,
Robert H. Svenson,
Stanley Yachnin
Publication year - 1969
Publication title -
proceedings of the national academy of sciences of the united states of america
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 5.011
H-Index - 771
eISSN - 1091-6490
pISSN - 0027-8424
DOI - 10.1073/pnas.63.2.334
Subject(s) - sephadex , biochemistry , phytohemagglutinins , fucose , phaseolus , mannose , glucosamine , glycoprotein , biology , hemagglutination , size exclusion chromatography , chemistry , antibody , botany , immune system , t cell , lymphocyte activation , immunology , enzyme
Commercially obtained phytohemagglutinin (PHAP) derived fromPhaseolus vulgaris contains 17 different protein bands when analyzed by acrylamide gel electrophoresis. When it is subjected to CM-Sephadex chromatography followed by molecular sieving on Sephadex G150, several species of potent mitogenic proteins, which differ greatly in their hemagglutinating capacity, are obtained. A low hemagglutinating mitogen (L-PHAP), homogeneous by several different criteria, is the most potent mitogen isolated, and also possesses potent leukoagglutinating activity. It is a glycoprotein with a molecular weight of 115,000, containing glucosamine, mannose, xylose, and fucose or arabinose. Also isolated is a mixture of at least two closely related proteins possessing high hemagglutinating capacity, with hemagglutination titers 250 times more potent than L-PHAP. This material is a slightly less potent mitogen than L-PHAP and also possesses leukoagglutinating capacity, although of a lower order of magnitude. Its amino acid and carbohydrate composition are similar to L-PHAP, but it contains approximately twice as much carbohydrate and is slightly larger as determined by molecular sieving.