title: Intact protein electrospray ionization tandem mass spectrometry can be the sole technique used for confirming the structure of a variant hemoglobin.
abstract: A mutation within the human alpha-globin resulting in an 18 u mass increment was characterized by sequencing the intact M(r) 15000 protein in the gas phase by electrospray ionization tandem mass spectrometry. No separation of the variant protein from its normal counterpart and other components of the hemolysate was performed prior to mass spectrometric analysis. Collision-induced dissociation of the globin molecules carrying 18 protons was affected by their exposure to methane in the collision cell of a triple quadrupole mass spectrometer equipped with an electrospray ion source. Consideration of the major fragmentation ions (b758+, y"284+ and y"617+) narrowed down the mutation site to the last 28 amino acid residues within the protein sequence. Examination of low-abundance product ions allowed the unequivocal identification of the mutation as alpha 136 Leu --> Met, a hemoglobin previously named Hb Chicago.
authors.affiliation: Children's Hospital Oakland Research Institute, California 94609-1809, USA.
authors.forename: H E
authors.initials: HE
authors.lastname: Witkowska
authors.forename: B N
authors.initials: BN
authors.lastname: Green
authors.forename: M
authors.initials: M
authors.lastname: Morris
authors.forename: C H
authors.initials: CH
authors.lastname: Shackleton
mesh_terms: D000595:Amino Acid Sequence; D000596:Amino Acids / Q000032:analysis; D005914:Globins / Q000737:chemistry; D006453:Hemoglobinopathies / Q000097:blood; D006455:Hemoglobins, Abnormal / Q000737:chemistry*; D006801:Humans; D007223:Infant; D013058:Mass Spectrometry; D008969:Molecular Sequence Data
chemical_list: D000596:Amino Acids; D006455:Hemoglobins, Abnormal; C050262:hemoglobin Chicago; D005914:Globins