Renaturated ricin toxin B chain made in Escherichia coli is soluble, stable, and biologically active.
Escherichia coli cells transformed with plasmids containing ricin B-chain coding sequences are shown to express this heterologous protein in inclusion bodies. After denaturation and renaturation of the product in the presence of glutathione and lactose, the recombinant ricin B-chain is soluble, biologically active and stable. Cytotoxicity of heterodimer with this protein and ricin A-chain is bound to be only ten times less than of native ricin. Recombinant B-chain alone was nontoxic to cells (ID50 > 10(-6)M). Our data suggest that N-glycosylation of ricin B-chain is not required for its biological activity.
Tonevitsky
A
A
Institute for Genetics of Microorganisms, Moscow, Russia.
Toptygin
A
A
Agapov
I
I
Pfueller
U
U
Frankel
A
A
D002460:Cell Line; D002470:Cell Survival / Q000187:drug effects*; D002850:Chromatography, Gel; D004591:Electrophoresis, Polyacrylamide Gel; D004797:Enzyme-Linked Immunosorbent Assay; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism; D005978:Glutathione / Q000737:chemistry; D007785:Lactose / Q000737:chemistry; D010957:Plasmids; D011489:Protein Denaturation; D011994:Recombinant Proteins / Q000737:chemistry / Q000633:toxicity; D012276:Ricin / Q000096:biosynthesis / Q000737:chemistry* / Q000302:isolation & purification / Q000633:toxicity*; D012995:Solubility
D011994:Recombinant Proteins; D012276:Ricin; D005978:Glutathione; D007785:Lactose