Capillary zone electrophoresis and mass spectrometry for the characterization of genetic variants of human hemoglobin.
This paper describes a simple and rapid analytical method for the structural identification of abnormal human hemoglobins. Globin chains obtained by precipitation of erythrocyte hemolysate in cold acetone are directly analyzed by capillary zone electrophoresis in coated capillaries without any prior treatment. The speed and the high resolving power of capillary zone electrophoresis allow fast differentiation of hemoglobins with similar charges. Capillary zone electrophoretic tryptic mapping has also been performed for each globin, so that complete variant characterization can be achieved by direct comparison of the variant tryptic map with the corresponding normal one. Coupling electrophoretic data with analysis of enzymatic digests by mass spectrometry according to the "fast atom bombardment mapping" procedure makes it possible to quickly identify amino acid variations. This paper describes how the method can be applied to the characterization of common and uncommon variants and underlines the advantages and limitations of the procedure along with its potential uses in structural analysis of proteins.
Ferranti
P
P
ICMIB/CNR Naples, Italy.
Malorni
A
A
Pucci
P
P
Fanali
S
S
Nardi
A
A
Ossicini
L
L
D002851:Chromatography, High Pressure Liquid; D004586:Electrophoresis / Q000379:methods; D014644:Genetic Variation; D005914:Globins / Q000032:analysis; D006454:Hemoglobins / Q000032:analysis* / Q000235:genetics; D006455:Hemoglobins, Abnormal / Q000032:analysis; D006801:Humans; D010449:Peptide Mapping; D012822:Silicon Dioxide; D016339:Spectrometry, Mass, Fast Atom Bombardment / Q000379:methods; D014357:Trypsin / Q000378:metabolism
D006454:Hemoglobins; D006455:Hemoglobins, Abnormal; D012822:Silicon Dioxide; D005914:Globins; D014357:Trypsin