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The PH-20 protein in human spermatozoa
K Sabeur1, G N Cherr, A I Yudin
1Department of Obstetrics and Gynecology, University of California, Davis 95616, USA.
Journal of Andrology
|March 1, 1997
Summary
PH-20, a sperm protein, exhibits hyaluronidase activity in human sperm, with two forms identified. This suggests PH-20 plays a key role in human fertilization.
Area of Science:
- Reproductive Biology
- Biochemistry
- Cell Biology
Background:
- PH-20 is a sperm plasma-membrane protein with known hyaluronidase activity in various mammals.
- Its specific role and characteristics in human sperm require detailed investigation.
Purpose of the Study:
- To characterize the PH-20 protein in noncapacitated and capacitated human sperm.
- To determine the hyaluronidase activity of PH-20 at different pH levels and identify its localization within human sperm.
Main Methods:
- Immunoblotting using SDS-PAGE and polyclonal antibodies to detect PH-20 forms.
- Hyaluronic acid substrate gel analysis and microplate assays to measure hyaluronidase activity.
- Transmission electron microscopy and immunogold labeling for ultrastructural localization.
Main Results:
- Two forms of PH-20 (64 kDa and 53 kDa) were identified in human sperm.
- Both forms exhibited hyaluronidase activity, with the 64-kDa form active at neutral and acid pH, and the 53-kDa form active only at acid pH.
- PH-20 was localized to the plasma membrane of the sperm head and the inner acrosomal membrane after the acrosome reaction.
Conclusions:
- PH-20 possesses hyaluronidase activity in human sperm.
- The biochemical properties and localization suggest PH-20 is the human sperm hyaluronidase, crucial for fertilization.