Purification and characterization of bolaffinine, a toxic protein from Boletus affinis Peck (Boletaceae).
A toxic protein of Mr 22,000, called bolaffinine, has been purified from the mushroom, Boletus affinis Peck (Boletaceae), with a 0.65% yield using a procedure involving four steps: ammonium sulfate precipitation, two chromatographies on ion-exchange columns and gel filtration on Sephadex G-75. It is thermolabile and can be inactivated by organic solvents. Its isoelectric point lies between pH 9 and 10. It contains 234 amino acids, no free SH groups and one disulfide bridge as evidenced by reactions with 5-5'-dithiobis-nitrobenzoate (DTNB) and 2-mercaptoethanol. When injected into mice, it provokes the death of animals within 16-24 h. The 24 h LD50 is 61 mg/kg of body weight. Bolaffinine inhibits protein synthesis in hepatoma tissue culture and in a rabbit reticulocyte lysate.
D000596:Amino Acids / Q000032:analysis; D000818:Animals; D001487:Basidiomycota / Q000032:analysis*; D001681:Biological Assay; D055598:Chemical Phenomena; D011232:Chemical Precipitation; D002627:Chemistry, Physical; D002845:Chromatography; D005656:Fungal Proteins / Q000302:isolation & purification* / Q000494:pharmacology / Q000633:toxicity; D007928:Lethal Dose 50; D008114:Liver Neoplasms, Experimental / Q000378:metabolism; D008297:Male; D051379:Mice; D009183:Mycotoxins; D014176:Protein Biosynthesis; D011817:Rabbits; D012156:Reticulocytes / Q000187:drug effects / Q000378:metabolism
D000596:Amino Acids; D005656:Fungal Proteins; D009183:Mycotoxins; C051088:Bolaffinine protein, Boletus affinis Peck