title: Determination of endogenous peptides in the porcine brain: possible construction of peptidome, a fact database for endogenous peptides.
abstract: Peptides play crucial roles in many physiological events. However, a database for endogenous peptides has not yet been developed, because the peptides are easily degraded by proteolytic enzymes during extraction and purification. In this study, we demonstrated that the data for endogenous peptides could be collected by minimizing the proteolytic degradation. We separated porcine brain peptides into 5250 fractions by 2-dimensional chromatography (first ion-exchange and second reversed-phase high-performance liquid chromatography), and 75 fractions of average peptide contents were analyzed in detail by mass spectrometers and a protein sequencer. Based on the analysis data obtained in this study, more than 10000 peptides were deduced to be detected, and more than 1000 peptides to be identified starting from 2 g of brain tissue. Thus, we deduce that it is possible to construct a database for endogenous peptides starting from a gram level of tissue by using 2-dimensional high-performance liquid chromatography coupled with a mass spectrometer.
authors.affiliation: Department of Pharmacology, National Cardiovascular Center Research Institute, 5-7-1 Fujishirodai, Suita, Osaka 565-8565, Japan. minamino@ri.ncvc.go.jp
authors.forename: Naoto
authors.initials: N
authors.lastname: Minamino
authors.forename: Junko
authors.initials: J
authors.lastname: Tanaka
authors.forename: Hiromiki
authors.initials: H
authors.lastname: Kuwahara
authors.forename: Takahiro
authors.initials: T
authors.lastname: Kihara
authors.forename: Yoshinori
authors.initials: Y
authors.lastname: Satomi
authors.forename: Masami
authors.initials: M
authors.lastname: Matsubae
authors.forename: Toshifumi
authors.initials: T
authors.lastname: Takao
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D001923:Brain Chemistry; D002851:Chromatography, High Pressure Liquid / Q000379:methods; D030562:Databases, Protein; D013058:Mass Spectrometry / Q000379:methods; D008969:Molecular Sequence Data; D010455:Peptides / Q000032:analysis* / Q000737:chemistry; D013552:Swine
chemical_list: D010455:Peptides