The sperm-specific proteins of the edible oyster (European flat oyster (Ostrea edulis)) are products of proteolytic processing

Extraction of sperm proteins from the bivalve mollusc Ostrea edulis shows them to contain a normal complement of core histones, together with three sperm-specific proteins, OE1 and OE2, plus the shorter OE3, which shows substantial microheterogeneity. OE1 and OE2 have a very similar amino acid composition, cyanogen bromide (CNBr) cleavage yields products of identical size and possesses a trypsin-resistant core peptide, together indicating that they are closely homologous histone H1-like proteins. Western blotting shows that OE1 and OE2 are closely related to the histone H1-like protein PL-II* of Mytilus trossulus. The amino acid composition of OE3 shows it to be a protamine-like PL-IV type protein. Edman degradation of a CNBr peptide from OE2 gave the sequence (M)KAAFAKGLKSGALVRPKGS-which has 85% identity to a sequence located towards the C-terminal end of the globular domain of the PL-II* protein of M. trossulus. An O. edulis sperm cDNA library yielded a clone of 428 bp. A genomic clone including an open reading frame (ORF) of 750 bp was isolated by PCR amplification from genomic DNA. Hypothetical translation showed the ORF to encode OE1 (or OE2) immediately followed by OE3, separated by a proteolytic processing site. This arrangement (a two-protein ORF) is also found in M. trossulus and Ensis minor.