Profiling of neuropeptides using gradient reversed-phase high-performance liquid chromatography with novel detection methodologies.
Biological tissues and fluids are subjected to gradient reversed-phase high-performance liquid chromatography (RP-HPLC) separation and the neuropeptide profile of the collected fractions is obtained by radioreceptor assay (RRA) using a broad-based competing ligand. Radioimmunoassay (RIA) is also used to detect specific neuropeptides in the HPLC-purified fractions. Further confirmation of the identity of the peptides present in the tissue is obtained by mass spectrometry (MS) in the fast atom bombardment (FAB) mode. FAB-MS produces the protonated molecular ion of the peptide and allows direct measurement of underivatized peptides at the nanogram level, with increased molecular specificity. FAB-MS-MS identifies a unique amino acid sequence-determining ion in the mass spectrum of a peptide and offers maximum molecular specificity. This analytical chromatography methodology is applied to the study of the molecular basis of several disease states by monitoring several peptidergic pathways and individual peptides and their metabolic relationships. Molecular mechanisms involved in pain, stress, tumor formation, and neurological studies are studied.
D000818:Animals; D002851:Chromatography, High Pressure Liquid; D004194:Disease / Q000378:metabolism; D004700:Endocrine System Diseases / Q000378:metabolism; D006801:Humans; D013058:Mass Spectrometry; D009369:Neoplasms / Q000378:metabolism; D009419:Nerve Tissue Proteins / Q000032:analysis*; D009619:Nociceptors / Q000502:physiology
D009419:Nerve Tissue Proteins