CAUDA EPIDIDYMAL SPERM INTERACTIONS WITH SEMINAL-VESICLE FLUID

The interaction of rat cauda epididymal sperm cAMP-dependent protein kinase (PKA) with seminal vesicle fluid (SVF) proteins was examined. Specific proteins in SVF act as substrates for the sperm cell PKA. The apparent molecular weights of these proteins are 45.0, 31.5, 17.2, 14.7, and 13.3 kDa. The phosphorylation of one low-molecular-weight cauda sperm protein is blocked in the presence of SVF. There is no PKA enzyme activity in SVF. The presence of phosphate transfer activity between the sperm cell enzyme and the SVF proteins is species dependent. For example, mouse and rat SVF proteins are efficient phosphate acceptors, but there is no phosphorylation activity when hamster SVF is used as the enzyme substrate. The sperm cell samples were also assessed for membrane integrity. Specifically, cauda sperm cells used in these assays were judged to be intact when examined microscopically using the fluorescent vital dye carboxyfluorodiacetate. Although there was enzyme activity in the supernatants of the rat sperm cell samples, in the protein kinase assay it required three times as much supernatant volume (compared with intact cell sample volume) to measure the activity. Supernatant enzyme activity did not increase with washing, indicating that the cells were not damaged by this procedure. The enzyme itself does not adhere to the sperm cells, so the PKA enzyme activity is most likely oriented on the external surface of the sperm cell.