title: High-throughput method for N-terminal sequencing of proteins by MALDI mass spectrometry.
abstract: A high-throughput method for sequencing of N termini of proteins by using postsource decay (PSD) of matrix-assisted laser desorption/ionization mass spectrometry has been developed. After a protein blotted on the PVDF membrane was successively reduced, S-alkylated, and guanidinated, its N-amino group was coupled to biotinylcysteic acid. The protein was then extracted from the membrane and digested with trypsin. The derivatized N-terminal fragment was then specifically isolated from the tryptic digest with avidin resins, and its de novo sequencing was successfully performed by PSD utilizing a sulfonic acid group introduced to the N terminus.
authors.affiliation: Life Science Laboratory, Shimadzu Corporation, Kyoto 604-8511, Japan, Department of Chemistry, Nara Women's University, Nara 630-8506, Japan. yamagu@shimadzu.co.jp
authors.forename: Minoru
authors.initials: M
authors.lastname: Yamaguchi
authors.forename: Takashi
authors.initials: T
authors.lastname: Nakazawa
authors.forename: Hiroki
authors.initials: H
authors.lastname: Kuyama
authors.forename: Takashi
authors.initials: T
authors.lastname: Obama
authors.forename: Eiji
authors.initials: E
authors.lastname: Ando
authors.forename: Taka-aki
authors.initials: TA
authors.lastname: Okamura
authors.forename: Norikazu
authors.initials: N
authors.lastname: Ueyama
authors.forename: Shigemi
authors.initials: S
authors.lastname: Norioka
mesh_terms: D000818:Animals; D001710:Biotin / Q000031:analogs & derivatives* / Q000737:chemistry; D003544:Cysteic Acid / Q000031:analogs & derivatives* / Q000737:chemistry; D006801:Humans; D007797:Laminin / Q000737:chemistry; D009113:Muramidase / Q000737:chemistry; D009842:Oligopeptides / Q000737:chemistry; D011506:Proteins / Q000737:chemistry*; D020539:Sequence Analysis, Protein / Q000379:methods*; D012710:Serum Albumin, Bovine / Q000737:chemistry; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization / Q000379:methods*
chemical_list: D007797:Laminin; D009842:Oligopeptides; D011506:Proteins; C516561:biotinylcysteic acid; D012710:Serum Albumin, Bovine; D001710:Biotin; D003544:Cysteic Acid; D009113:Muramidase