Striped mullet (Mugil cephalus) hemoglobin system: multiplicity and functional properties.
The most frequent (90%) phenotype of the hemoglobin system of M. cephalus presented two major hemoglobins, the more anodal HbI accounting for approximately 70% of the total. The two hemoglobin components separated by ion-exchange chromatography were analyzed by reverse-phase HPLC and electrospray ionization-mass spectrometry which revealed a more complex pattern: HbI consists in four different globins, two β (named β1 and β3) and two co-eluting α chains (α1 and α2); HbII consists in three globins, one β chain (named β2) and the same α1 and α2 present in HbI. The oxygen-binding properties of both hemoglobin components purified by DEAE cellulose were almost identical to those of the hemolysate: stripped hemoglobin showed a large Bohr effect which was enhanced by chloride ions and, at a larger extent, by organic phosphates which, at acidic pH values gave rise to the Root effect. A series of oxygen-binding experiments at increasing GTP concentrations was carried out in order to compare GTP-binding activities in the absence and presence of physiological amounts of chloride. The results indicated that hemoglobin do have two sites for GTP binding. In the absence of chloride, the two sites cannot be discriminated, whereas in the presence of chloride, a competition between the two anions occurred for both GTP-binding sites. The presence of multiple hemoglobin components with identical properties confirms that hemoglobin heterogeneity that often occurs in fish cannot be only explained as an evolutionary response to the physiological and/or environmental needs of the species.
Department of Sciences Applied to Biosystems, University of Cagliari, Cittadella Universitaria, 09042, Monserrato (CA), Italy.
Alessandra
A
Olianas
Claudia
C
Meloni
Irene
I
Messana
Maria T
MT
Sanna
Massimo
M
Castagnola
Barbara
B
Manconi
Susanna
S
Salvadori
Bruno
B
Giardina
Mariagiuseppina
M
Pellegrini
D000818:Animals; D002712:Chlorides / Q000378:metabolism; D002851:Chromatography, High Pressure Liquid; D002852:Chromatography, Ion Exchange; D003636:DEAE-Cellulose; D006160:Guanosine Triphosphate / Q000378:metabolism; D055554:Hemoglobin Subunits / Q000737:chemistry* / Q000302:isolation & purification / Q000378:metabolism*; D006863:Hydrogen-Ion Concentration; D010100:Oxygen / Q000378:metabolism; D023701:Smegmamorpha / Q000097:blood*; D021241:Spectrometry, Mass, Electrospray Ionization
D002712:Chlorides; D055554:Hemoglobin Subunits; D006160:Guanosine Triphosphate; D003636:DEAE-Cellulose; D010100:Oxygen