Several synthetic endothelin (ET) analogues of the C-terminal ET hexapeptide (ET16-21) were analyzed by radio-receptor competition binding assays and biologic activity using both ETA and ETB receptor subtypes. In addition, we produced a hybridoma monoclonal antibody, anti-ET15-21, that appeared to crossreact with the entire ET molecule and was able to neutralize its biologic activity. Antibody binding was measured with competition enzyme-linked immunosorbent assays and a surface plasmon resonance-based biosensor (BIA technology). The ET16-21 moiety was modified with systematic replacement of each residue by alanine (Ala-scan). Whereas the C-terminal residues (Asp(18), Ile(20), and particularly Trp(21)) were very important for both receptor binding and immunologic activity, Ala substitution in positions 16, 17, and 19 hardly affected such activities. Analysis of another series of synthetic ET16-21 analogues with the His(16) residue replaced by a non-amino-acidic block confirmed that the last two C-terminal residues are essential for receptor and antibody binding, whereas the central region of this hexapeptide is much more tolerant to modification. However, a critical steric conformation of the active hexapeptide is necessary.
|Autori interni:||MAZZONI, MARIA ROSA|
|Autori:||Rovero P; Galoppini C; Laricchia-Robbio L; Mazzoni M; Revoltella R|
|Titolo:||Structure-activity analysis of C-terminal endothelin analogues|
|Anno del prodotto:||1998|
|Digital Object Identifier (DOI):||10.1097/00005344-199800001-00071|
|Appare nelle tipologie:||1.1 Articolo in rivista|