Binding of mannose-6-phosphate and heparin by boar seminal plasma PSP-II, a member of the spermadhesin protein family.

PSP-I/PSP-II, a heterodimer of glycosylated spermadhesins, is the major component of boar seminal plasma. Similarly to other spermadhesins, the PSP-II subunit is a lectin which displays heparin- and zona pellucida glycoprotein-binding activities. We have investigated the ligand binding capabilities of the heterodimer and the isolated subunits using several polysaccharides, glycoproteins, and phospholipids. PSP-II binds the sulfated polysaccharides heparin and fucoidan in a dose-dependent and seemingly-specific manner. In addition, PSP-II binds oligosaccharides containing exposed mannose-6-phosphate monoester groups and the binding is selectively inhibited by mannose-6-phosphate and glucose-6-phosphate. Inhibition experiments indicate that binding of PSP-II to sulfated polysaccharides and mannose-6-phosphate-containing oligosaccharides involves distinct but possibly overlapping binding sites. Heterodimer formation with PSP-I abolishes both the heparin and the mannose-6-phosphate binding capabilities, suggesting that the corresponding sites may be located at the dimer interface. Using the crystal structure of PSP-I/PSP-II heterodimer as a template, we have explored possible binding sites which satisfy the observed binding characteristics. In the proposed models, PSP-II Arg43 appears to play a pivotal role in both heparin- and mannose-6-phosphate-complexation as well as in heterodimer formation.