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c-Abl proto-oncoprotein is expressed and tyrosine phosphorylated in human sperm cell
1Department of Obstetrics and Gynecology, Medical College of Ohio, Toledo 43614, USA. rnaz@gemini.mco.edu
Abstract:
The presence and possible role of c-Abl proto-oncoprotein was investigated in human sperm cell. The c-Abl monoclonal antibody (mAb), against the protein tyrosine kinase domain of v-Abl protein, reacted specifically with the acrosomal region of methanol-fixed capacitated and non-capacitated human sperm cell in the indirect immunofluorescence technique. The c-Abl mAb predominantly recognized two protein bands of 145 kD and 95 kD in detergent-solubilized (Triton X-100 and NP-40) sperm and testes preparations in the Western blot procedure. The 95 kD protein band reacted stronger than the 145 kD band and was the only band detected in the lithium diiodosalicylate (LIS)-solubilized sperm preparation, and even in the Triton X-100/NP-40 extracts of sperm of some men. In the in vitro kinase assay using the Triton X-100-solubilized capacitated sperm preparation, the 95 kD protein was autophosphorylated at the tyrosine residues, which was inhibited in the presence of c-Abl mAb. The tyrosine phosphorylation of sperm proteins, especially of the 95 kD protein, has been shown to have a vital role in human sperm function, namely, the sperm capacitation/acrosomal exocytosis and binding to zona pellucida of oocyte. These findings suggest that the c-Abl or c-Abl-like proteins are present in mature sperm cells that are tyrosine autophosphorylated and may have a role in human sperm cell function.
Insights
The c-Abl proto-oncoprotein is present in human sperm, with a 95 kD form showing tyrosine autophosphorylation. This protein may play a crucial role in human sperm function, including capacitation and oocyte binding.
Area of Science:
- Cell Biology
- Reproductive Biology
- Molecular Biology
Background:
- The c-Abl proto-oncoprotein's role in human sperm function is largely unknown.
- Protein tyrosine phosphorylation is critical for sperm capacitation and acrosomal exocytosis.
Purpose of the Study:
- To investigate the presence and potential function of c-Abl in human sperm.
- To identify the specific c-Abl protein forms and their enzymatic activity within sperm.
Main Methods:
- Indirect immunofluorescence using c-Abl monoclonal antibody (mAb).
- Western blot analysis of sperm and testes preparations.
- In vitro kinase assay to assess protein autophosphorylation.
Main Results:
- c-Abl mAb specifically recognized the acrosomal region of human sperm.
- Western blots identified 145 kD and 95 kD c-Abl protein bands, with the 95 kD band being predominant.
- The 95 kD protein exhibited tyrosine autophosphorylation, inhibited by the c-Abl mAb.
Conclusions:
- c-Abl or c-Abl-like proteins are present in mature human sperm.
- These proteins undergo tyrosine autophosphorylation, suggesting a role in sperm function.
- The findings implicate c-Abl in critical processes like sperm capacitation and oocyte binding.