A mutational analysis of receptor binding sites of interleukin-1 beta: differences in binding of human interleukin-1 beta muteins to human and mouse receptors
| Autor: | E. Edelmann, John P. Priestle, Markus G. Grütter, U. Feige, U. Joss, K. Vosbeck, Albert Schmitz, J. Van Oostrum |
|---|---|
| Rok vydání: | 1994 |
| Předmět: |
Models
Molecular Circular dichroism DNA Mutational Analysis Molecular Sequence Data Bioengineering Biology Crystallography X-Ray Biochemistry Mice Structure-Activity Relationship Escherichia coli Animals Humans Amino Acid Sequence Receptor Beta (finance) Site-directed mutagenesis Molecular Biology Genetics Binding Sites Molecular Structure Circular Dichroism Mutagenesis Receptors Interleukin-1 Biological activity Molecular biology Affinities Cell culture Mutagenesis Site-Directed Biotechnology Interleukin-1 |
| Zdroj: | Protein engineering. 7(5) |
| ISSN: | 0269-2139 |
| Popis: | The 3-D crystal structure of interleukin-1 beta (IL-1 beta) has been used to define its receptor binding surface by mutational analysis. The surface of IL-1 beta was probed by site-directed mutagenesis. A total of 27 different IL-1 beta muteins were constructed, purified and analyzed. Receptor binding measurements on mouse and human cell lines were performed to identify receptor affinities. IL-1 beta muteins with modified receptor affinity were evaluated for structural integrity by CD spectroscopy or X-ray crystallography. Changes in six surface loops, as well as in the C- and N-termini, yielded muteins with lower binding affinities. Two muteins with intact binding affinities showed 10- to 100-fold reduced biological activity. The surface region involved in receptor binding constitutes a discontinuous area of approximately 1000 A2 formed by discontinuous polypeptide chain stretches. Based on these results, a subdivision into two distinct local areas is proposed. Differences in receptor binding affinities for human and mouse receptors have been observed for some muteins, but not for wild-type IL-1 beta. This is the first time a difference in binding affinity of IL-1 beta muteins to human and mouse receptors has been demonstrated. |
| Databáze: | OpenAIRE |
| Externí odkaz: |
|