Detection of phosphopeptides in bovine adrenal medulla by using iron (III) immobilized-affinity chromatography and mass spectrometry

Eur. J. Mass Spectrom. 5, 279–284 (1999)
DOI: 10.1255/ejms.286

Detection of phosphopeptides in bovine adrenal medulla by using iron (III) immobilized-affinity chromatography and mass spectrometry

Shihua Li and Chhabil Dass^*
Department of Chemistry, The University of Memphis, Campus Box 526060, Memphis, TN 38152, USA. E-mail: cdass@cc.memphis.edu

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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