Isolation and characterization of the hemoglobin from the lanceolate fluke Dicrocoelium dendriticum.

The hemoglobin of the flatworm Dicrocoelium dendriticum, a lanceolate fluke which infests the hepatic ducts of certain mammals, has been isolated by gel filtration and ion-exchange chromatography. The molecular weight of the denatured protein was found to be 15500, a value in the same range as hemoglobin subunits. The fact that the native hemoglobin has an apparent molecular weight of 22000 in 0.01 M phosphate buffer, pH 7.4, suggests limited aggregation. The protein contains, as all other myoglobins and hemoglobins, one molecule of non-covalently associated ferroprotoporphyrin IX per polypeptide chain. It forms the same ligand derivatives with very similar spectral properties as vertebrate hemoglobins. The high oxygen affinity (p50 is 0.07--0.1 mmHg or 9.3--13.3 Pa at 20 degrees C and pH 7.0) and the absence of heme-heme interaction of (Hill coefficient nH=1.0) are properties which this heme protein shares with other monomeric hemoglobins from invertebrate and lower vertebrate organisms. The native hemoglobin exists in two forms, having isoelectric points of 4.51 and 4.53, which do not differ in their amino-acid compositions. Dansylation indicated that the amino-terminal amino-acid residue is alanine. The carboxy-terminal sequence, determined by carboxypeptidase A digestion of the globin, is -His-Ala-Leu.