Implication of the oligomeric state of the N-terminal PTX3 domain in cumulus matrix assembly

Elena Ievoli, Ragnar Lindstedt, Antonio Inforzato, Antonella Camaioni, Francesca Palone, Anthony J. Day, Alberto Mantovani, Giovanni Salvatori, Antonietta Salustri

Research output: Contribution to journalArticle

Abstract

Pentraxin 3 (PTX3) plays a key role in the formation of the hyaluronan-rich matrix of the cumulus oophorus surrounding ovulated eggs that is required for successful fertilization and female fertility. PTX3 is a multimeric protein consisting of eight identical protomers held together by a combination of non-covalent interactions and disulfide bonds. Recent findings suggest that the oligomeric status of PTX3 is important for stabilizing the cumulus matrix. Because the role of PTX3 in the cumulus resides in the unique N-terminal sequence of the protomer, we investigated further this issue by testing the ability of distinct Cys/Ser mutants of recombinant N-terminal region of PTX3 (N _PTX3) with different oligomeric arrangement to promote in vitro normal expansion in cumuli from Ptx3-null mice. Here we report that the dimer of the N _PTX3 is unable to rescue cumulus matrix organization, and that the tetrameric assembly of the protein is the minimal oligomeric state required for accomplishing this function. We have previously demonstrated that PTX3 binds to HCs of IαI and TSG-6, which are essential for cumulus matrix formation and able to interact with hyaluronan. Interestingly, here we show by solid-phase binding experiments that the dimer of the N _PTX3 retains the ability to bind to both IαI and TSG-6, suggesting that the octameric structure of PTX3 provides multiple binding sites for each of these ligands. These findings support the hypothesis that PTX3 contributes to cumulus matrix organization by cross-linking HA polymers through interactions with multiple HCs of IαI and/or TSG-6. The N-terminal PTX3 tetrameric oligomerization was recently reported to be also required for recognition and inhibition of FGF2. Given that this growth factor has been detected in the mammalian preovulatory follicle, we wondered whether FGF2 negatively influences cumulus expansion and PTX3 may also serve in vivo to antagonize its activity. We found that a molar excess of FGF2, above PTX3 binding capacity, does not affect in vitro cumulus matrix formation thus ruling out this possibility. In conclusion, the data strength the view that PTX3 acts as a nodal molecule in cross-linking HA in the matrix.

Original languageEnglish
Pages (from-to)330-337
Number of pages8
JournalMatrix Biology
Volume30
Issue number5-6
DOIs
Publication statusPublished - Jun 2011

Keywords

  • Cumulus matrix assembly
  • FGF2
  • Hyaluronan
  • IαI
  • Pentraxin3
  • TSG-6

ASJC Scopus subject areas

  • Molecular Biology

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    Ievoli, E., Lindstedt, R., Inforzato, A., Camaioni, A., Palone, F., Day, A. J., Mantovani, A., Salvatori, G., & Salustri, A. (2011). Implication of the oligomeric state of the N-terminal PTX3 domain in cumulus matrix assembly. Matrix Biology, 30(5-6), 330-337. https://doi.org/10.1016/j.matbio.2011.05.002