Affinity sites for N-acetyl-beta-D-glucosaminidase on the surface of rat epididymal spermatozoa.

The binding of N-acetyl-beta-D-glucosaminidase from rat epididymal fluid to the surface of spermatozoa from the cauda epididymis was measured in the presence of sugars, its phosphorylated derivatives, or after treatment of the cells or the enzyme with agents that alter the integrity of proteins or carbohydrates. The binding was saturable, with a Kd in the nanomolar range, was inhibited with phosphorylated derivates of fructose, and did not depend on Ca2+, showing that it is different from the mannose 6-P-recognizing system existing in other tissues for this and other acid hydrolases. Treatment of the cells with sodium periodate or trypsin inhibited the binding, showing that a glycoprotein of the plasmalemma is involved in the affinity site. Fructose or phosphorylated derivates were not detected in the proteins of the epididymal fluid with HPLC. However, with the method used, the presence of these compounds cannot be ruled out, if among the proteins of the fluid there are only a small number of acid hydrolases containing this sugar.