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Construction and Characterization of Ciliary Neurotrophic Factor (CNTF) Antagonists: Microenvironmental Difference in the CNTF Receptor Between Rat and Chicken Cells for Recognizing the D1 Cap Region
Author(s) -
Inoue Makoto,
Karita Hiromi,
Nakayama Chikao,
Noguchi Hiroshi
Publication year - 1997
Publication title -
journal of neurochemistry
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.75
H-Index - 229
eISSN - 1471-4159
pISSN - 0022-3042
DOI - 10.1046/j.1471-4159.1997.69010095.x
Subject(s) - ciliary neurotrophic factor , receptor , mutant , microbiology and biotechnology , leukemia inhibitory factor receptor , biology , dorsal root ganglion , neurotrophic factors , medicine , chemistry , anatomy , biochemistry , dorsum , embryonic stem cell , gene , leukemia inhibitory factor
Antagonistic mutants of ciliary neurotrophic factor (CNTF) were constructed and their properties characterized. K155A and K155W mutants lost cell survival promoting activity for chicken dorsal root ganglion (DRG) neurons and inhibited the activity of the wild type. However, they retained slight agonistic activity for the survival of rat DRG neurons, indicating there is a difference between chicken and rat cells for receptor recognition around the D1 cap region including K155 residue. The chicken receptor recognizes the D1 cap region more strictly than does the rat receptor. The substitution of F152, which locates at the top of the D1 cap region, was combined with the K155A mutation. A combination of the two mutations gave an antagonistic feature to not only chicken but also rat cells. Both F152S/K155A and F152D/K155A mutants lacked cell survival promoting activity and had an antagonistic effect on rat DRG neurons. The three‐dimensional structure of CNTF suggests the following. F152 and K155 bind to the receptor with hydrophobic and electrostatic interactions, respectively. F152 locates close to L156 with a van der Waals contact, and K155 contacts with Q42 through a hydrogen bond. Both interactions play indispensable roles in maintaining the structure around the D1 cap region of CNTF.