Asymmetric hemoglobins, their thiol content, and blood glutathione of the scalloped hammerhead shark, Sphyrna lewini.
Starch gel electrophoresis pH 8.6, or PAGE pH 8.9, of the scalloped hammerhead shark hemolysates showed three hemoglobins (Hb). An additional Hb between the two most mobile electrophoretic components was seen in starch gel electrophoresis, pH 8.1, and also in highly loaded PAGE gels. The relative concentration of these Hbs was variable among individuals, when accessed at pH 8.1. Dilution of hemolysates led to a redistribution of the Hb tetramer subunits. Under denaturing conditions, the unfractionated hemolysate was resolved in 3 Hb subunits. Isolated Hbs, named SL I-SL IV, showed unusual subunit compositions: SL I, the least mobile, is "b3c"; SL II is "a2bc"; SL III and SL IV are composed only by "a" subunits. Hemoglobins in the whole hemolysate have an average of two reactive cysteines per tetramer, which were not easily S-thiolated by glutathione, as is the case for related species. After hemoglobin denaturation, six additional -SH groups were titrated by Ellman's reagent. Methemoglobin content was low in the erythrocytes of nine examined specimens, 1.13 +/- 1.90%. High values for total erythrocyte glutathione (GSH) were found: 4.5 +/- 0.7 mM; n = 7. The ratio of 1.4 +/- 0.4 GSH/Hb is higher than usually reported for mammalians.
Dafré
A L
AL
Depto. de Biofísica, UFRGS, Porto Alegre, Brazil. ALDAFRE@CCB.USFC.BR
Reischl
E
E
D000818:Animals; D004586:Electrophoresis / Q000379:methods; D004912:Erythrocytes / Q000737:chemistry; D005978:Glutathione / Q000031:analogs & derivatives / Q000097:blood* / Q000737:chemistry; D019803:Glutathione Disulfide; D006400:Hematocrit; D006454:Hemoglobins / Q000737:chemistry*; D006863:Hydrogen-Ion Concentration; D008706:Methemoglobin / Q000032:analysis / Q000737:chemistry; D012754:Sharks / Q000097:blood*; D013213:Starch; D013438:Sulfhydryl Compounds / Q000032:analysis* / Q000737:chemistry
D006454:Hemoglobins; D013438:Sulfhydryl Compounds; D013213:Starch; D008706:Methemoglobin; D005978:Glutathione; D019803:Glutathione Disulfide