Caractérisation des isoformes de la protéine SPAM1 (Sperm Adhesion Molecule 1) et identification de ses partenaires d'interactions dans les spermatozoïdes
|Abstract:||Sperm Adhesion Molecule 1 (SPAM1) is a sperm protein that has a hyaluronidase activity in its N-terminus, aiding in the dispersal of the cumulus cells surrounding the egg. It also has a zona pellucida (ZP) binding activity in its C-terminus. Our previous studies showed that there are two potential SPAM1 isoforms that have a molecular weight of ~70 and 80 kDa in the bovine species. From these studies, we hypothesized that these two SPAM1 isoforms had different C-terminal domains, different origins (testis or epididymis) and were localised in the acrosomal or post-acrosomal regions of spermatozoa. Seeing as it is the C-terminal domain that is involved in ZP binding, we aimed to characterize the two C-terminal domains in order to better understand SPAM1’s role in gamete interactions. Although the 3’ nucleotide sequences were identical, two Spam1 transcripts varying by the presence or absence of 90 nucleotides (exon 3 of the Spam1 gene) were found in both testicular and epididymal tissues. During our studies, we also identified PH-20, a potential SPAM1 homolog. In order to determine if PH-20 is one of the two potential SPAM1 isoforms that is recognized by our antibody directed against the N-terminal domain, we attempted the production of a PH-20 recombinant protein, without success. We also sought to determine if SPAM1 is part of a multimeric protein complex involved in spermatozoa-ZP interactions, as reported in humans. Our results suggest that SPAM1 is associated with AKAPs, which are anchoring proteins abundantly found in the fibrous sheath of sperm flagella. Characterizing the SPAM1 isoforms, its homolog PH-20, as well as the multimeric protein complexes SPAM1 is part of, are important in order to better understand SPAM1’s role in gamete interactions.|
|Document Type:||Mémoire de maîtrise|
|Open Access Date:||24 April 2018|
|Collection:||Thèses et mémoires|
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