Purification and properties of blood hemoglobin from the fresh-water cladocera, Moina macrocopa and Daphnia magna

Abstract

1. 1. From extracts of the blood of the pink animals of Moina macrocopa and Daphnia magna, hemoglobins were purified by sucrose density gradient centrifugation, gel filtration, and DEAE-cellulose column chromatography. 2. 2. Sedimentation and electrophoretic analyses indicated that the hemoglobins were homogeneous. 3. 3. The molecular properties of Moina hemoglobin were as follows: sedimentation coefficient 17.8 S; diffusion coefficient 2.62 · 10-7 cm2/sec; intrinsic viscosity 0.045 dl/g; mol. wt. 6.70 · 105; frictional ratio 1.31; isoelectric point 5.2; iron content 0.317%; and heme content 37-38. 4. 4. The values of P50 were 2.1 mm Hg for Moina and 3.5 mm Hg for Daphnia at 20° and pH 7.2, and the Bohr effect was hardly observed in both species. 5. 5. Absorption spectra showed a similarity with mammalian hemoglobin with regard to spectral maxima and relative extinction coefficients. © 1971.