Fig. 3. Relative binding of ¹²⁵I-mZP2 and ¹²⁵I-mZP3 glycoproteins to bacterial expressed mouse proacrosin peptide. (A) Coomassie Blue stained proteins separated by reducing SDS-PAGE after extraction from inclusion bodies. Lane 1, pre-induction; lane 2, post-induction showing expression of a major 32 kDa peptide (arrow). (B) Binding of unfractionated ¹²⁵I-mZP glycoproteins to expressed 32 kDa peptide on western blots. Lane 3, pre-induced bacterial proteins probed with ¹²⁵I-mZP glycoproteins; lane 4, post-induction showing binding of ¹²⁵I-mZP glycoproteins to the 32 kDa peptide. (C) Binding of purified ¹²⁵I-mZP2 (lane 5) and purified ¹²⁵I-mZP3 (lane 6) to the 32 kDa peptide. (D) Immunological identification of the 32 kDa peptide as expressed residues 38-288 of proacrosin. Lane 7, immune serum. Lane 8, pre-immune serum. (E) Autoradiograph of purified ¹²⁵I-mZP glycoproteins separated by SDS-PAGE. Lane 9, unfractionated ¹²⁵I-mZP glycoproteins. Lane 10, purified ¹²⁵I-mZP2 equivalent to 120 kDa. Lane 11, purified ¹²⁵I-mZP3 equivalent to 83 kDa. (F) Relative binding of purified ¹²⁵I-mZP2 and ¹²⁵I-mZP3 to expressed proacrosin 32 kDa peptide. All experiments were repeated at least 3 times and results shown are representative of the data as a whole. See text for details.

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