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A spectrofluorometric investigation, using 1-anilino-naphthalene-8-sulphonate, of the interaction between washed bovine spermatozoa and seminal plasma or egg-yolk lipoprotein

Summary. The dye 1-anilino-naphthalene-8-sulphonate (ANS) bound at 94·5 ± 2·7 pmol per 10⁶ spermatozoa (K_D = 1·83 ± 0·13 x 10^–5 M) and 7·30 ± 0·17 pmol per mg seminal plasma protein (K_D = 2·19 ± 0·06 x 10^–5 M). Equilibration of excess ANS with a mixture of washed spermatozoa and seminal plasma resulted in a significant increase in fluorescence over that calculated by summation of the individual levels (P < 0·05). A less pronounced but significant increase was seen when an egg-yolk lipoprotein, previously shown to have cryoprotective properties, was similarly added to washed spermatozoa (P < 0·05). This increase in fluorescence was not reduced by washing spermatozoa–lipoprotein mixtures, suggesting that an interaction between the lipoprotein and the cell membrane had occurred.

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