Immunological dissimilarity in protein component (dynein 1) between outer and inner arms within sea urchin sperm axonemes

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Abstract
The 0.5 M KCl-treatment solubilizes the outer arms from sea urchin sperm axonemes. Almost 30% of A-polypeptide, corresponding to dynein 1 in sodium dodecylsulfate-polyacrylamide gel, was solubilized by this treatment (as SEA-dynein 1). EM observation indicated that the extracted axonemes lacked the outer arms in various degrees. The SEA-dynein 1 was purified and an antiserum against it was prepared in rabbits. The specificity of antiserum to dynein 1 was determined by immunoelectrophoresis and Ouchterlony''s double-diffusion test. The anti-dynein 1 serum inhibited ATPase activity of purified SEA-dynein 1 by 95%. By the indirect peroxidase-conjugated antibody method, the loci of SEA-dynein 1 within the intact, salt-extracted and mechanically disrupted axonemes were determined to be the outer arms: deposition of electron-dense materials which represents their localization was detected at the distal ends of the outer arms, in the case of intact axonemes. The 5-6 cross-bridge was hardly decorated. No decoration was seen in the salt-extracted axonemes lacking all the outer arms. In disrupted axonemes, which consist of single to several peripheral doublets, electron-dense materials were deposited only at the outer arms. Axonemal ATPase activity sensitive to antiserum [73%] was solubilized by repeated salt-extractions. One-half of A-polypeptide (SEA-dynein 1 located at the outer arms) was contained in the pooled extracts. The extracted axonemes contained another half of A-polypeptide (SUA-dynein 1 supposed to locate at the inner arms) and retained 31% of axonemal ATPase activity that was almost resistant to antiserum. Solubilized SUA-dynein 1 was immunologically the same as SEA-dynein 1. In situ SUA-dynein 1 probably did not receive anti-dynein 1 antibodies, coinciding with the result obtained for salt-extracted axonemes lacking all the outer arms by the enzyme-antibody method mentioned above. These observations suggest the immunological dissimilarity in dynein 1 between outer and inner arms but do not tell us that the inner arms do not contain dynein 1.