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Abstract

Immobilized iron (III) displays binding affinity toward phosphate groups of phosphopeptides/proteins. This interaction has been exploited to selectively isolate phosphopeptides from the bovine adrenal medulla gland. The peptide-rich fraction was first obtained from the homogenized tissue by solid-phase extraction using a C-18 cartridge. Phosphopeptides were selectively isolated from this fraction by iron (III) immobilized-metal affinity chromatography [Fe(III)-IMAC] and characterized by high-performance liquid chromatography/electrospray ionization and matrix-assisted laser desorption/ionization mass spectrometry techniques. Further corroborative evidence of the presence of phosphopeptides was obtained by treating the Fe(III)-IMAC-retained fraction with acid phosphatase.

Keywords

Phosphopeptides adrenal medulla affinity chromatography LC/ESI-MS MALDI-TOF-MS

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Detection of Phosphopeptides in Bovine Adrenal Medulla by Using Iron (III) Immobilized-Affinity Chromatography and Mass Spectrometry

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