Site-Specific Conjugation to Interleukin 4 Containing Mutated Cysteine Residues Produces Interleukin 4-Toxin Conjugates with Improved Binding and Activity
作者： Robert J. Kreitman , Raj K. Puri , Pamela Leland , Byungkook Lee , Ira Pastan
DOI: 10.1021/BI00204A027
关键词:
摘要: Fusion of a ligand to another protein frequently impairs the binding ligand. Recombinant toxins composed mutants Pseudomonas exotoxin (PE) fused C-terminus human interleukin 4 (IL4) are cytotoxic IL4 receptor- (IL4R-) bearing tumor cells but bind IL4R with only 1% affinity IL4. We have developed method connect toxin which allows junction be moved location on would minimize impairment. designed in residue 28, 38, 68, 70, 97, or 105 was substituted cysteine. All purified bound 60-100% IL4, indicating that structure essentially unchanged. The were then each conjugated through disulfide bond PE35, truncated form PE contains single PE35 at 10-fold improved and more than recombinant IL4-toxin is position 129 containing 97 had lower activity. These results indicate ligand-protein can selectively enhance conjugate effectiveness, implications could made regarding regions important for binding.
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