A structural study on elcatonin, a novel synthetic analogue of eel calcitonin, by fast atom bombardment and tandem mass spectrometry.
Elcatonin (ELC), a synthetic analogue of eel calcitonin, successfully used in the treatment of diseases characterized by an increase of osteoclastic activity, has been fully sequenced by combined enzymatic hydrolysis and fast atom bombardment (FAB) mass spectrometric methodology. The FAB mass spectrometric analysis on the entire molecule gave only the cluster corresponding to the molecular ion. Digestion with trypsin afforded four oligopeptides corresponding to fragments 1-11, 12-18, 19-24 and 25-32. B/E daughter ion analysis performed in turn on each protonated molecule ion in the tryptic mixture allowed complete sequencing of fragments 1-11 and 12-18, while in fragments 19-24 and 25-32 the portions 23-24 and 25-26 remained respectively unclarified. Investigation on the single oligopeptide isolated by preparative high-performance liquid chromatography and chymotryptic digestion of the molecule failed to provide any new information. One step of Edman degradation on tryptic digest permitted attribution of the 25-26 sequence to Thr-Asp. The connectivity between tryptic ELC fragments and the 23-24 sequence were proved by cleavage with V8 protease, which gave the oligopeptides 1-15 and 16-32. These were exhaustively analysed by FAB mass spectrometry.
Pasqualucci
C
C
ISF Laboratories for Biomedical Research, Milan, Italy.
Visconti
M
M
Danieli
B
B
Rubino
F M
FM
D002116:Calcitonin / Q000031:analogs & derivatives* / Q000032:analysis / Q000302:isolation & purification; D002851:Chromatography, High Pressure Liquid; D013058:Mass Spectrometry; D008970:Molecular Weight; D009842:Oligopeptides / Q000032:analysis; D016339:Spectrometry, Mass, Fast Atom Bombardment
D009842:Oligopeptides; D002116:Calcitonin; C012755:elcatonin