title: Sequence of the human erythrocyte phosphoglycerate mutase by microsequencer and mass spectrometry.
abstract: We have previously reported the isolation in pure form of the human erythrocyte phosphoglycerate mutase isozyme B. We now report the sequence of the whole protein and the identification of its N-terminal blocking group. The protein tryptic peptides of phosphoglycerate mutase isozyme B were isolated by high performance liquid chromatography and their sequence determined by microsequencing. The sequence and the nature of the blocking group of the N-terminal tryptic peptide was shown to be N-acetyl-Ala-Ala-Tyr-Lys by mass spectrometry. Overlaps of the tryptic peptides were obtained by studying the V8 Staphylococcus aureus protease peptides of the aminoethylated phosphoglycerate mutase isozyme B either by microsequencing or by mass spectrometry. The procedure used allowed us to obtain the sequence on a very small amount of material and in a short period of time. Our data agree well with those derived from the cDNA nucleotide sequence described by Sakoda et al. (Sakoda, S., Shanske, S., DiMauro, S., and Schon, E. A. (1988) J. Biol. Chem. 263, 16899-16905). In addition, our data directly indicate that the initiation codon does not introduce a methionine as N-terminal amino acid and allowed the identification of the acetyl N-terminal group.
authors.affiliation: Institut National de la Santé et de la Recherche Médicale U.91, Hôpital Henri Mondor, Créteil, France.
authors.forename: Y
authors.initials: Y
authors.lastname: Blouquit
authors.forename: M C
authors.initials: MC
authors.lastname: Calvin
authors.forename: R
authors.initials: R
authors.lastname: Rosa
authors.forename: D
authors.initials: D
authors.lastname: Promé
authors.forename: J C
authors.initials: JC
authors.lastname: Promé
authors.forename: F
authors.initials: F
authors.lastname: Pratbernou
authors.forename: M
authors.initials: M
authors.lastname: Cohen-Solal
authors.forename: J
authors.initials: J
authors.lastname: Rosa
mesh_terms: D000595:Amino Acid Sequence; D001731:Bisphosphoglycerate Mutase / Q000032:analysis*; D002851:Chromatography, High Pressure Liquid; D004912:Erythrocytes / Q000201:enzymology*; D006801:Humans; D013058:Mass Spectrometry; D008969:Molecular Sequence Data; D010446:Peptide Fragments / Q000032:analysis; D010770:Phosphotransferases / Q000032:analysis*
chemical_list: D010446:Peptide Fragments; D010770:Phosphotransferases; D001731:Bisphosphoglycerate Mutase