Characterization of bioactive peptides in bovine adrenal medulla by a combination of fast HPLC and ESI-MS.
A method based upon a combination of fast high-performance liquid chromatography (HPLC) and electrospray ionization (ESI)-mass spectrometry (MS) is developed for the analysis of bioactive peptides in bovine adrenal medulla. The fast HPLC uses a short column (33 mm x 4.6 mm) packed with nonporous silica-based C-18 stationary phase. Prior to HPLC separation, the medulla was homogenized and the peptide-rich fraction was isolated from it by solid-phase extraction. In-source collision-induced dissociation and tandem MS were used to obtain the sequence of the suspected peptides. Several peptides, including Met-Enk, Leu-Enk, Leu-Enk-Lys, bovine adrenal medullary (BAM)-12 (Met-Enk-RRVGRPE), Leu-Enk-Arg, and YGGT, were unambiguously identified. The first four peptides are the products of proenkephalin A precursor protein and Leu-Enk-Arg belongs to the dynorphin family and is derived from proenkephalin B (prodynorphin) precursor. The database search revealed that YGGT is a part of the sequence of five different precursor proteins.
Wang
Haiping
H
Department of Chemistry, The University of Memphis, Campus Box 526060, Memphis, TN 38152, USA.
Dass
Chhabil
C
D000313:Adrenal Medulla / Q000378:metabolism*; D000818:Animals; D002417:Cattle; D002851:Chromatography, High Pressure Liquid; D010455:Peptides / Q000032:analysis* / Q000378:metabolism; D021241:Spectrometry, Mass, Electrospray Ionization
D010455:Peptides