[Production of biologically active recombinant ricin B-chain].
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 containing this protein and ricin A-chain is found to be only ten times lower, than that of native ricin. Recombinant B-chain alone was nontoxic to cells (ID50 > 10(-6) M). Our data suggest that ricin B-chain oligosaccharides are essential for stability preserving protein from proteolytic degradation in cells.
Tonevitskiĭ
A G
AG
Institute for Phytochemistry, University of Witten/Herdecke, Witten, Germany.
Toptygin
A Iu
AIu
Agapov
I I
II
Rakhmanova
V A
VA
Shamshiev
A T
AT
Alekseev
Iu O
IuO
Pfueller
U
U
Frankel
A
A
D001483:Base Sequence; D002470:Cell Survival / Q000187:drug effects; D003001:Cloning, Molecular; D004926:Escherichia coli / Q000235:genetics; D006868:Hydrolysis; D008969:Molecular Sequence Data; D009838:Oligodeoxyribonucleotides; D011489:Protein Denaturation; D011994:Recombinant Proteins / Q000737:chemistry / Q000235:genetics / Q000494:pharmacology; D012276:Ricin / Q000737:chemistry / Q000235:genetics / Q000494:pharmacology*
D009838:Oligodeoxyribonucleotides; D011994:Recombinant Proteins; D012276:Ricin
Poluchenie biologicheski aktivnoĭ rekombinantnoĭ B-tsepi ritsina.