| Abstract: | Previous studies have identified an "actin-like" protein in human sperm by indirect immunofluorescence microscopy with various probes, but no real biochemical confirmation of actin was made. In this study, two-dimensional (2-D) gels of Nonidet P-40 (NP-40) extracts of purified human sperm cells revealed a protein with appropriate pI and molecular weight coordinates for actin. When excised from a 2-D gel, cleaved with N-chlorosuccinimide, and separated on a sodium dodecyl sulfate slab gel, this putative actin showed a cleavage pattern identical to those from known actin. Furthermore, when sperm were immediately purified from whole semen and extracted with 0.3% NP-40, actin was detected almost entirely in the soluble fraction, indicative of unpolymerized actin. Conclusions from these experiments support those implied by direct immunofluorescence microscopy: actin is present in human sperm and appears to be mainly unpolymerized. |
