Purification and properties of different forms of modeccin, the toxin of Adenia digitata. Separation of subunits with inhibitory and lectin activity.
1. The subunits were isolated of modeccin (subsequently referred to as modeccin 4B), the toxin purified from the roots of Adenia digitata by affinity chromatography on Sepharose 4B [Gasperi-Campani, Barbieri, Lorenzoni, Montanaro, Sperti, Bonetti & Stirpe (1978) Biochem J. 174, 491-496]. They are an A subunit (mol.wt. 26 000), which inhibits protein synthesis, and a B subunit (mol.wt. 31 000), which binds to cells. Both sununits, as well as intact modeccin, gave single bands on sodium dodecyl sulphate/polyacrylamide-gel electrophoresis, but showed some heterogeneity on isoelectric focusing and on polyacrylamide-gel electrophoresis at pH 9.5. 2. A second form of modeccin, not retained by Sepharose 4B, was purified by affinity chromatography on acid-treated Sepharose 6B: this form is subsequently termed modeccin 6B 3. Modeccin 6B has a molecular weight indistinguishable from that of modeccin 4B, and consists of two subunits of mol.wts. 27 000 and 31 000, joined by a disulphide bond. The subunits were not isolated because of their high insolubility in the absence of sodium dodecyl sulphate. 4. As compared with modeccin 4B, modeccin 6B is slightly less toxic to animals, does not agglutinate erythrocytes, and is a more potent inhibitor of protein synthesis in a lysate of rabbit reticulocytes, giving 50% inhibition at the concentration of 0.31 microgram/ml.
D000818:Animals; D055598:Chemical Phenomena; D002621:Chemistry; D002846:Chromatography, Affinity; D003864:Depression, Chemical; D004591:Electrophoresis, Polyacrylamide Gel; D005033:Ethylmaleimide; D006384:Hemagglutination; D066298:In Vitro Techniques; D037102:Lectins / Q000302:isolation & purification* / Q000494:pharmacology; D008970:Molecular Weight; D037121:Plant Lectins; D010947:Plants, Toxic / Q000032:analysis*; D014176:Protein Biosynthesis; D011817:Rabbits; D051381:Rats; D012156:Reticulocytes / Q000187:drug effects / Q000378:metabolism; D054790:Ribosome Inactivating Proteins, Type 2; D014118:Toxins, Biological / Q000302:isolation & purification* / Q000494:pharmacology / Q000633:toxicity
D037102:Lectins; D037121:Plant Lectins; D054790:Ribosome Inactivating Proteins, Type 2; D014118:Toxins, Biological; C016004:modeccin; D005033:Ethylmaleimide