[Heterogeneity of rat hemoglobin. Isolation of constituent globin chains by HPLC and their amino acid sequence analysis]

In order to elucidate the molecular mechanism of the heterogeneity of rat hemoglobin, hemoglobin components in adult rat hemolysate were isolated and differences in the primary structures of their constituent globins were determined. The results obtained were as follows: 1) Crystalline hemoglobin prepared from the hemolysate of an adult male Wistar rat was separated into nine hemoglobin components by means of DEAE-cellulose column chromatography, each giving a single band on polyacrylamide gel electrophoresis. 2) Of these hemoglobins, six major components, designated DE-I, II, IV, V, VI and VII, were further subjected to reverse-phase HPLC to yield their respective constituent alpha and beta globins. 3) Peptide mapping and NH2-terminal amino acid sequence analysis of the beta globins confirmed that DE-I and II hemoglobins possessed the previously known III beta globin, while DE-IV, V, VI and VII hemoglobins contained the previously known II beta globin. 4) The mechanism underlying the heterogeneity of rat hemoglobin was discussed on the basis of these findings.