Mutants of interleukin 13 with altered reactivity toward interleukin 13 receptors.
Interleukin 13 (IL13) belongs to a family of cytokines whose members exhibit structural homology, despite amino acid sequence dissimilarity. For example, while of limited sequence homology, IL13 and IL4 share a signaling receptor, IL13/4 receptor, on a variety of human normal cells. However, a subclass of IL4-independent IL13 receptors is overexpressed on certain transformed cells, including human malignant gliomas. We introduced mutations into human (h) IL13 to determine the site(s) involved in interaction with the shared receptor and/or the glioma-associated receptor. This analysis identified at least three protein regions that are needed for signaling through the shared receptor. These regions were localized to alpha-helices A, C, and D and were mainly separate from the region(s) needed to interact with the glioma-associated receptor. Glutamic acids at positions 13 and 16 in hIL13 alpha-helix A, arginine and serine at positions 66 and 69 in helix C, and arginine at position 109 in helix D were found to be important in inducing biological signaling since their specific mutation resulted in loss and/or gain of function phenomena. We demonstrate that the molecular requirements of hIL13 to interact with its respective receptors are generally distinct and can be controlled by mutagenesis of the cytokine.
Thompson
J P
JP
Section of Neurosurgery/H110, Department of Surgery, Pennsylvania State University College of Medicine, Hershey, Pennsylvania 17033-0850, USA.
Debinski
W
W
D001483:Base Sequence; D002455:Cell Division; D002470:Cell Survival / Q000187:drug effects; D002478:Cells, Cultured; D002942:Circular Dichroism; D003603:Cytotoxins / Q000494:pharmacology; D017931:DNA Primers; D005909:Glioblastoma / Q000378:metabolism / Q000473:pathology; D006801:Humans; D018793:Interleukin-13 / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism*; D053661:Interleukin-13 Receptor alpha1 Subunit; D016297:Mutagenesis, Site-Directed; D018123:Receptors, Interleukin / Q000378:metabolism*; D053660:Receptors, Interleukin-13; D011994:Recombinant Proteins / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism; D014407:Tumor Cells, Cultured; D019010:Vascular Cell Adhesion Molecule-1 / Q000096:biosynthesis
D003603:Cytotoxins; D017931:DNA Primers; C508181:IL13RA1 protein, human; D018793:Interleukin-13; D053661:Interleukin-13 Receptor alpha1 Subunit; D018123:Receptors, Interleukin; D053660:Receptors, Interleukin-13; D011994:Recombinant Proteins; D019010:Vascular Cell Adhesion Molecule-1