. The Biological bulletin. Biology; Zoology; Biology; Marine Biology. 176 T. L. VANDERGON ET AL. Table II Oxygen binding characteristics of metacercaria hemoglobin PH P50(mmHg) Hill constant ±* ± ± ± ± ± ± ± 10 7 6 1 SD. ance of the highly vascularized metanephridial tissue; however, transmission electron micrographs of gut ceca and gut lining cells revealed no evidence of material that resembled the characteristic annelid high molecular weight extracellular hemoglobins. Figure 3 shows the elution profil
. The Biological bulletin. Biology; Zoology; Biology; Marine Biology. 176 T. L. VANDERGON ET AL. Table II Oxygen binding characteristics of metacercaria hemoglobin PH P50(mmHg) Hill constant ±* ± ± ± ± ± ± ± 10 7 6 1 SD. ance of the highly vascularized metanephridial tissue; however, transmission electron micrographs of gut ceca and gut lining cells revealed no evidence of material that resembled the characteristic annelid high molecular weight extracellular hemoglobins. Figure 3 shows the elution profile of metacercaria he- moglobin from HPLC gel filtration which separates mol- ecules on the basis of their size. Two fractions were eluted; fraction A eluted in the void volume of the col- umn (MW > 500,000 daltons) and fraction B eluted at 16,000 daltons. Standard gel filtration on a Sephacryl S- 400 column (Fig. 4) indicated that fraction A has a mo- lecular weight of x 106 daltons. HPLC ion exchange chromatograms of whole meta- cercaria lysate and gel filtration fractions of metacercaria hemoglobin are shown in Figure 5a-c. Ion exchange sep- aration of whole metacercaria hemoglobin resulted in four distinct protein components (Fig. 5a). Metacercaria gel filtration fraction A (MW ~ X 106 daltons) when chromatographed on the ion exchange column demonstrated one broad peak with a distinct shoulder. Table III Comparison of oxygen binding characteristics of metacercaria and Amphitrite ornata hemoglobins Bohr P50(mm Hg) Hill constant coefficient A. ornata coelomic cell in cellulo (SD, ) (SD, ) — (pH ) cell suspension* — (pH ) in vitro* NS (pH ) vascular* NS (pH ) vascular** 3= _ (pH ) Metacercaria (pH ) If) r-l ^r LU u z < m a. o iS) m < rr < O LU t/5- () LU mui o w. * Data is from Mangum el ai. (1975). ** Da
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