Diphtheria toxin receptor binding domain substitution with interleukin-2: genetic construction and properties of a diphtheria toxin-related interleukin-2 fusion protein.
We have genetically replaced the diphtheria toxin receptor binding domain with a synthetic gene encoding interleukin-2 (IL-2) and a translational stop signal. The diphtheria toxin-related T-cell growth factor fusion gene encodes a 70 586-d polypeptide, pro-IL-2-toxin. The mature form of IL-2-toxin has a deduced mol. wt of 68,086 and is shown to be exported to the periplasmic compartment of Escherichia coli (pABI508), and contain immunologic determinants intrinsic to both its diphtheria toxin and IL-2 components. IL-2-toxin has been purified from periplasmic extracts of recombinant strains of E. coli (pABI508) by immunoaffinity chromatography using immobilized anti-IL-2. The purified chimeric toxin is shown to selectively inhibit protein synthesis in IL-2 receptor bearing targeted cells, whereas cell lines which do not express the IL-2 receptor are resistant to IL-2-toxin action.
Williams
D P
DP
Evans Department of Clinical Research, University Hospital, Boston University Medical Center, MA 02118.
Parker
K
K
Bacha
P
P
Bishai
W
W
Borowski
M
M
Genbauffe
F
F
Strom
T B
TB
Murphy
J R
JR
D001426:Bacterial Proteins / Q000302:isolation & purification; D002460:Cell Line; D004247:DNA / Q000302:isolation & purification; D004167:Diphtheria Toxin; D004926:Escherichia coli / Q000235:genetics; D005813:Genes, Synthetic; D066257:Heparin-binding EGF-like Growth Factor; D015151:Immunoblotting; D036341:Intercellular Signaling Peptides and Proteins; D007376:Interleukin-2 / Q000235:genetics* / Q000378:metabolism; D010957:Plasmids; D011956:Receptors, Cell Surface; D011950:Receptors, Cholinergic / Q000235:genetics* / Q000378:metabolism; D011993:Recombinant Fusion Proteins / Q000235:genetics* / Q000378:metabolism; D011994:Recombinant Proteins / Q000235:genetics*
D001426:Bacterial Proteins; D004167:Diphtheria Toxin; D066257:Heparin-binding EGF-like Growth Factor; D036341:Intercellular Signaling Peptides and Proteins; D007376:Interleukin-2; D011956:Receptors, Cell Surface; D011950:Receptors, Cholinergic; D011993:Recombinant Fusion Proteins; D011994:Recombinant Proteins; D004247:DNA