Peptide quantification by tandem mass spectrometry.

Abstract

This manuscript reviews the literature on the mass spectrometry (MS) and tandem mass spectrometry (MS/MS) quantification of biologically important peptides that have been extracted from tissues. The most important aspect of this quantification process is the use of MS/MS to link the protonated molecule ion, (M + H)(+) , of the peptide with one or more of its amino acid sequence-determining fragment ions. The actual name of a peptide cannot be used in any study until the amino acid sequence of that peptide has been firmly established. This article reviews the analytical data obtained from the measurement of opioid peptides in human pituitary tissues. For example, the proopiomelanocortin (POMC)-derived beta-endorphin (BE) and the proenkephalin-derived methionine enkephalin (ME) opioid peptides have been quantified. The biogenesis of opioid neuropeptides is briefly reviewed; critical aspects of pituitary neuropeptides are discussed, including their localization and regulation, and their role in tumor formation; other analytical methods used to detect and measure neuropeptides are mentioned, including radioimmunoassay (RIA), radioreceptorassay (RRA), in situ hybridization, mRNA, and cDNA methods; and the MS and MS/MS methods are described. The use of stable isotope-incorporated synthetic peptide internal standards is described. Data are presented on the measurement of BE and ME in control pituitaries and in pituitary tumors (PRL-secreting and nonsecreting tumors). A significant alteration in the POMC peptide BE was found between the control and tumor tissues. That difference suggests that the POMC neuropeptidergic system had been down-regulated in those tumors. © 1997 John Wiley & Sons, Inc.