title: Domains of rat interleukin 1 beta involved in type I receptor binding.
abstract: In the present study the inhibitory effects of a panel of 21 monoclonal antibodies (moabs) to rat interleukin 1 beta (rIL-1 beta) on the binding of 125I-labeled rIL-1 beta to murine type I IL-1 receptors on EL4 cells were investigated. Furthermore, the epitopes of these moabs were determined by the use of the pepscan technique, and these epitopes were visualized on a three-dimensional model of rIL-1 beta. Some moabs (SILK 3, 4, 5, 6, and 22) inhibited receptor binding of radioiodinated rIL-1 beta at concentrations that are similar to the dissociation constant values of antibody-rIL-1 beta binding. Another group of moabs (SILK 7, 11, 20, 21, and 23) also inhibited receptor binding but only at concentrations that are 10-150 times higher than their dissociation constants. A large group of moabs did not affect receptor binding in the concentration range tested, and two moabs enhanced the binding of rIL-1 beta to type I receptors. The result of pepscan analysis shows that the moabs bound to one or more of the amino acid sequences 35-49, 66-85, 78-97, 106-124, and 123-143 of mature rIL-1 beta. Modeling of rIL-1 beta shows that the binding domains of SILK 4, 5, 6, and 22 (sequence 123-143) is located at the closed end of the molecule, indicating that this part of rIL-1 beta harbors domains that are crucial for type I receptor binding. The binding domain of SILK 3 (sequence 66-85) is also located at this end of the molecule. In contrast, the binding domains of SILK 7, 11, 20, 21, and 23 (sequence 78-97) are located at the open end of the molecule, which is at the same face as the amino- and carboxy-terminals. The binding domain of SILK 16 (sequence 106-124) is positioned at the center of the molecule. It is concluded that the closed end of rIL-1 beta contains sequences that are crucial for its binding to type I receptors on murine EL4 cells. Because of the high concentrations of antibodies to residues 78-97 of rIL-1 beta that are needed to interfere with receptor binding, the importance of these domains in binding to type I receptors remains uncertain.
authors.affiliation: Research Institute Neurosciences Vrije University, Faculty of Medicine, Department of Pharmacology, Amsterdam, The Netherlands.
authors.forename: K
authors.initials: K
authors.lastname: Schotanus
authors.forename: G M
authors.initials: GM
authors.lastname: Holtkamp
authors.forename: R H
authors.initials: RH
authors.lastname: Meloen
authors.forename: W C
authors.initials: WC
authors.lastname: Puijk
authors.forename: F
authors.initials: F
authors.lastname: Berkenbosch
authors.forename: F J
authors.initials: FJ
authors.lastname: Tilders
mesh_terms: D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology; D002460:Cell Line; D000939:Epitopes; D007375:Interleukin-1 / Q000737:chemistry* / Q000276:immunology / Q000378:metabolism*; D051379:Mice; D008807:Mice, Inbred BALB C; D008958:Models, Molecular; D051381:Rats; D017472:Receptors, Interleukin-1 / Q000378:metabolism*; D011994:Recombinant Proteins
chemical_list: D000911:Antibodies, Monoclonal; D000939:Epitopes; D007375:Interleukin-1; D017472:Receptors, Interleukin-1; D011994:Recombinant Proteins