Hemocyanin from Dolabella auricularia

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Abstract
It was shown that hemocyanin from a gastropod, Dolabella auricularia was dissociated into smaller units by passing through a DEAE-cellulose column. The sedimentation coefficient of DEAE-dissociated hemocyanin fraction I, which was not adsorbed by the ion exchanger, was 15 to 19 S. Fraction I was converted by prolonged contact with the ion exchanger into fraction II, which was adsorbed by DEAE-cellulose. In fraction II, two components with sedimentation coefficients of 4 and 7 S were observed. By means of gel filtration the molecular weights of 4 and 7 S components were estimated to be 60,000–70,000 and 150,000–160,000, respectively. Re-association of DEAE-dissociated hemocyanin in the presence of 0.5 M NaCl or 0.01 M CaCl2 was investigated by means of ultracentrifugation and electron microscopy. In contrast with Dolabella hemocyanin dissociated by EDTA (1), aggregation of fraction I by CaCl2 was random and incomplete. Fraction II compponents were scarcely re-associated by the salts. Oxygen affinity of DEAE-dissociated hemocyanin was twice as large as that of native Dolabella hemocyanin, while n-value in Hill's equation showed only a slight decrease.

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