Analysis of human IL-2/IL-2 receptor beta chain interactions: monoclonal antibody H2-8 and new IL-2 mutants define the critical role of alpha helix-A of IL-2.

Asn, Asp --> Lys, Asp --> Leu, show a correlation between diminished affinity for IL-2 receptor and reduced bioactivity measured on TS1beta cells. Mutein Asp Arg lose affinity for IL-2R and bioactivity simultaneously. Furthermore, during the course of the study we have found that mutein Asp20 --> Leu is an IL-2 antagonist. The biological effects of mAb H2-8 and the properties of new mutants at positions 17 and 20 demonstrate that this region of alpha helix-A is involved in IL-2-IL-2R beta interactions. -->
Molecular Sequence: PDB 1ILM; 3INK
Substance Nomenclature: 0 (Antibodies, Monoclonal)
0 (Interleukin-2)
0 (Receptors, Interleukin-2)
30KYC7MIAI (Aspartic Acid)
9007-49-2 (DNA)
GMW67QNF9C (Leucine)
Entry Date(s): Date Created: 19970701 Date Completed: 19971010 Latest Revision: 20131121
Update Code: 20240829
DOI: 10.1006/cyto.1996.0192
PMID: 9237811
Autor: Eckenberg R; Immunogénétique Cellulaire, Institut Pasteur, Paris, France., Xu D, Moreau JL, Bossus M, Mazie JC, Tartar A, Liu XY, Alzari PM, Bertoglio J, Theze J
Jazyk: angličtina
Zdroj: Cytokine [Cytokine] 1997 Jul; Vol. 9 (7), pp. 488-98.
DOI: 10.1006/cyto.1996.0192
Abstrakt: Interleukin 2 (IL-2) interacts with a receptor (IL-2R) composed of three subunits (IL-2R alpha, IL-2R beta and IL-2R gamma). IL-2R beta plays a critical role in signal transduction. An anti-human IL-2 mAb (H2-8) produced after immunization with peptide 1-30 of IL-2 was found to recognize the region occupied by Asp20, at the exposed interface between alpha-helices A and C. Muteins at position 17 and 20 are not recognized by mAb H2-8. mAb H2-8 specifically inhibits the IL-2 proliferation of TS1beta cells which are dependent on the expression of human IL-2R beta chain for IL-2 proliferation. Substitution at internal position Leu17 demonstrates that this position is essential for IL-2 binding and IL-2 bioactivity. New IL-2 mutants at position Asp20 have been analysed. Substitutions Asp --> Asn, Asp --> Lys, Asp --> Leu, show a correlation between diminished affinity for IL-2 receptor and reduced bioactivity measured on TS1beta cells. Mutein Asp Arg lose affinity for IL-2R and bioactivity simultaneously. Furthermore, during the course of the study we have found that mutein Asp20 --> Leu is an IL-2 antagonist. The biological effects of mAb H2-8 and the properties of new mutants at positions 17 and 20 demonstrate that this region of alpha helix-A is involved in IL-2-IL-2R beta interactions.
Databáze: MEDLINE
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