Effects induced by hydroxyl radicals on salmon calcitonin: a RP-HPLC, CD and TEM study

The effects of hydroxyl radical attack on a peptidic drug were studied in vitro. Different chemico-physical techniques were used to investigate structural damage induced by oxidative stress conditions in salmon calcitonin (sCT), a peptide hormone used in treating osteoporosis. Reversed-phase liquid...

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Bibliographic Details
Published in:Biochimica et biophysica acta 2003-09, Vol.1623 (1), p.33-40
Main Authors: Gaudiano, Maria Cristina, Diociaiuti, Marco, Bertocchi, Paola, Valvo, Luisa
Format: Article
Language:English
Subjects:
Calcitonin - chemistry
Chromatography, High Pressure Liquid
Circular Dichroism
Crystallography - methods
Hydroxyl Radical - chemistry
Hydroxyl radicals
Macromolecular Substances
Microscopy, Electron
Oxidative stress
Protein Conformation
RP-HPLC
Salmon calcitonin
TEM
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Summary:The effects of hydroxyl radical attack on a peptidic drug were studied in vitro. Different chemico-physical techniques were used to investigate structural damage induced by oxidative stress conditions in salmon calcitonin (sCT), a peptide hormone used in treating osteoporosis. Reversed-phase liquid chromatography (RP-HPLC), circular dichroism (CD) and transmission electron microscopy (TEM) were applied to measure formation of oxidation/degradation products and to reveal the conformational and ultrastructural modifications in the presence of OH· free radicals. Hydroxyl radicals were obtained from ferrous sulfate and ascorbic acid mixtures. The RP-HPLC results revealed the formation of new chromatographic peaks indicating a number of degradation/oxidation products formed in the presence of OH· free radicals. CD spectra showed slight protein conformational modifications as well as aggregation. TEM confirmed sCT aggregation and suggested the formation of fibrillar aggregates.
ISSN:0304-4165
0006-3002
1872-8006
DOI:10.1016/S0304-4165(03)00158-2