title: Determination of neutral haemoglobin variants by immobilized pH gradient, reversed-phase high-performance liquid chromatography and fast-atom bombardment mass spectrometry: the case of a Hb Torino alpha 43 (CE1) Phe----Val.
abstract: A neutral haemoglobin variant was identified by the combined use of different analytical methods. Isoelectric focusing on immobilized pH gradients (under denaturing conditions) allowed to detect and alpha chain variant. HPLC of tryptic digest showed that its amino-acid variation resided on T alpha 6 peptide (position 41-56). In the fast-atom bombardment (FAB) positive mass spectrum of the tryptic digest, the protonated molecular ion of the T alpha 6 peptide occurred 48 mass units lower than the normal T alpha 6 fragment, corresponding to a Phe----Val substitution. To a partial sequence determination (aminopeptidase digestion of the T alpha 6 peptide, followed by amino acid determination and FAB-MS analysis of the digestion-generated mixture) the substitution appeared to be on Phe43 (CE1). This variant is already known as Hb Torino. The procedure here described proved to be fast and simple, and feasible whenever neutral variants are supposed to occur.
authors.affiliation: Istituto di Chimica, Università Cattolica del S. Cuore, Roma.
authors.forename: M
authors.initials: M
authors.lastname: Castagnola
authors.forename: M
authors.initials: M
authors.lastname: Dobosz
authors.forename: R
authors.initials: R
authors.lastname: Landolfi
authors.forename: V L
authors.initials: VL
authors.lastname: Pascali
authors.forename: F
authors.initials: F
authors.lastname: de Angelis
authors.forename: L
authors.initials: L
authors.lastname: Vettore
authors.forename: G
authors.initials: G
authors.lastname: Perona
mesh_terms: D002675:Child, Preschool; D002851:Chromatography, High Pressure Liquid; D005260:Female; D014644:Genetic Variation; D006454:Hemoglobins / Q000302:isolation & purification; D006455:Hemoglobins, Abnormal / Q000235:genetics* / Q000302:isolation & purification; D006801:Humans; D013058:Mass Spectrometry; D010446:Peptide Fragments / Q000032:analysis; D010649:Phenylalanine; D014633:Valine
chemical_list: D006454:Hemoglobins; D006455:Hemoglobins, Abnormal; D010446:Peptide Fragments; C046798:hemoglobin Torino; D010649:Phenylalanine; D014633:Valine