Boar spermadhesins are 12-14 kDa lectins which coat the entire acrosomal cap sperm head surface. The large amount of spermadhesins AQN-1, AQN-2, AQN-3, and AWN-1 present on in vitro capacitated spermatozoa (approximately 7 x 10(6) molecules of each spermadhesin per cell) suggested that they may bind to major component(s) of the sperm surface. We have investigated both the aggregation state of spermadhesins in seminal plasma using gel filtration chromatography, and their ability to bind to the major phospholipids of the boar sperm plasma membrane, i.e. phosphorylcholine and phosphorylethanolamine. The bulk (90%) of spermadhesins AQN-3 and AWN-1 were eluted as aggregated proteins (Mr > 50,000) with the void volume of a Sephadex G-50 column; the remaining 10% of the total amount of seminal plasma AWN-1 and AQN-3 were recovered, together with the whole amount of AQN-1 and AQN-2, in a fraction containing low-molecular-mass proteins (Mr 16,000-30,000). None spermadhesin of either gel-filtration fraction bound to a phosphorylcholine affinity matrix. On the other hand, low-molecular-mass (monomeric or dimeric) AQN-3 and AWN-1 were the only spermadhesins retained in a phosphorylethanolamine affinity column. Both AQN-3 and AWN-1 purified from seminal plasma by reverse-phase HPLC retained their lipid-binding capability. In addition, immobilization of AQN-3 and AWN-1 onto a phosphorylethanolamine matrix did not interfere with the ability of the proteins to bind bovine glycoprotein PDC-109, indicating that the structural determinants for the binding lipid and carbohydrates lay on different structural domains.(ABSTRACT TRUNCATED AT 250 WORDS)