The Multi-PDZ domain protein MUPP1 as a lipid raft-associated scaffolding protein controlling the acrosome reaction in mammalian spermatozoa.
The success of acrosomal exocytosis, a complex process with a variety of interrelated steps, relies on the coordinated interaction of participating signaling molecules. Since scaffolding proteins are known to spatially organize sequential signaling pathways, we examined whether the Multi-PDZ domain protein MUPP1, recently identified in mammalian spermatozoa, is functionally active in controlling acrosomal secretion in mammalian sperm cells. To address this question, permeabilized mouse sperm were loaded with inhibitory antibodies against MUPP1 as well as with a photosensitive Ca(2+) chelator which allows a controlled release of acrosomal Ca(2+). The results revealed that MUPP1 controls initial tethering and docking of the acrosomal vesicle, whereas syntaxin 2, a t-SNARE protein also expressed in the acrosomal cap of mammalian spermatozoa, appears to take part in the final process of acrosomal fusion. Interestingly, using immunogold electron microscopy, it was found that MUPP1 is detectable in the region of the periacrosomal membrane. Furthermore, in isolated detergent-insoluble glycolipid-enriched membrane domains from epididymal spermatozoa, MUPP1 was found to show a striking association with the Triton X-100 insoluble membrane fraction, which did not change significantly upon sperm capacitation or partial chemical extraction of cholesterol. This evidence points to a role of MUPP1 as a membrane raft-associated molecular organizer, and suggests that mammalian spermatozoa may use a scaffolding protein and distinct membrane subdomains to spatially organize components involved in the process of acrosomal exocytosis.
Ackermann
Frauke
F
Department of Pharmacology and Toxicology, Philipps-University Marburg, Germany.
Zitranski
Nele
N
Heydecke
Diana
D
Wilhelm
Beate
B
Gudermann
Thomas
T
Boekhoff
Ingrid
I
D000177:Acrosome / Q000187:drug effects; D020101:Acrosome Reaction / Q000502:physiology*; D000818:Animals; D000906:Antibodies / Q000494:pharmacology; D002118:Calcium / Q000494:pharmacology; D002352:Carrier Proteins / Q000737:chemistry* / Q000378:metabolism*; D002417:Cattle; D003902:Detergents / Q000494:pharmacology; D005089:Exocytosis / Q000187:drug effects; D047908:Intracellular Signaling Peptides and Proteins; D008297:Male; D021962:Membrane Microdomains / Q000378:metabolism*; D008565:Membrane Proteins; D051379:Mice; D017434:Protein Structure, Tertiary; D021381:Protein Transport / Q000187:drug effects; D051381:Rats; D013094:Spermatozoa / Q000378:metabolism*; D014020:Tissue Extracts / Q000378:metabolism
D000906:Antibodies; D002352:Carrier Proteins; D003902:Detergents; D047908:Intracellular Signaling Peptides and Proteins; D008565:Membrane Proteins; C491750:Mpdz protein, mouse; C491749:Mpdz protein, rat; D014020:Tissue Extracts; D002118:Calcium