A study of the cytochromes of Octopus vulgaris Lam

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Abstract
The animal was bled and perfused with sea water until no trace of hemocyanin was present in the perfusion fluid. The skinned mantle and tentacle muscles were collected and frozen at -20[degree] C before grinding in a meat grinder. The ground material was suspended in sea water at 3[degree] for 30 minutes and strained through cheesecloth. This procedure was repeated 5 times and then the water was squeezed through muslin and the material homogenized in phosphate buffer (0.1 [image], pH 7.3, 2.5 l/kg frozen muscle). The suspension was centrifuged at 2000 g, resuspended and centrifuged again. The pooled supernatants were centrifuged at 30,000 g for 1 hour and the sediment was washed and made up to 50 mg dry weight/ml in the phosphate buffer. Difference spectra of the particles show the presence of a cytochrome system composed of cytochromes a, a3, b and c. Enzymic oxidation of succinate and cytochrome oxidase activity were also demonstrated in these particles.