Pentraxin 3 (PTX3) and heavy chains (HCs) of inter-alpha-trypsin inhibitor (I alpha I) are essential for hyaluronan ( HA) organization within the extracellular matrix of the cumulus oophorus, which is critical for in vivo oocyte fertilization and female fertility. In this study, we examined the possibility that these molecules interact and cooperate in this function. We show that HCs and PTX3 colocalize in the cumulus matrix and coimmunoprecipitate from cumulus matrix extracts. Coimmunoprecipitation experiments and solid-phase binding assays performed with purified human I alpha I and recombinant PTX3 demonstrate that their interaction is direct and not mediated by other matrix components. PTX3 does not bind to I alpha I subcomponent bikunin and, accordingly, bikunin does not compete for the binding of PTX3 to I alpha I, indicating that PTX3 interacts with I alpha I subcomponent HC only. Recombinant PTX3-specific N-terminal region, but not the PTX3-pentraxin C-terminal domain, showed the same ability as full-length protein to bind to HCs and to enable HA organization and matrix formation by Ptx3(-/-) cumulus cell oocyte complexes cultured in vitro. Furthermore, a monoclonal antibody raised against PTX3 N terminus, which inhibits PTX3/I alpha I interaction, also prevents recombinant full-length PTX3 from restoring a normal phenotype to in vitro-cultured Ptx3(-/-) cumuli. These results indicate that PTX3 directly interacts with HCs of I alpha I and that such interaction is essential for organizing HA in the viscoelastic matrix of cumulus oophorus, highlighting a direct functional link between the two molecules.
Scarchilli, L., Camaioni, A., Bottazzi, B., Negri, V., Doni, A., Deban, L., et al. (2007). PTX3 interacts with inter-alpha-trypsin inhibitor - Implications for hyaluronan organization and cumulus oophorus expansion. THE JOURNAL OF BIOLOGICAL CHEMISTRY, 282(41), 30161-30170.
|Tipologia:||Articolo su rivista|
|Citazione:||Scarchilli, L., Camaioni, A., Bottazzi, B., Negri, V., Doni, A., Deban, L., et al. (2007). PTX3 interacts with inter-alpha-trypsin inhibitor - Implications for hyaluronan organization and cumulus oophorus expansion. THE JOURNAL OF BIOLOGICAL CHEMISTRY, 282(41), 30161-30170.|
|IF:||Con Impact Factor ISI|
|Settore Scientifico Disciplinare:||Settore BIO/17|
|Revisione (peer review):||Sì, ma tipo non specificato|
|Digital Object Identifier (DOI):||http://dx.doi.org/10.1074/jbc.M703738200|
|Stato di pubblicazione:||Pubblicato|
|Data di pubblicazione:||2007|
|Titolo:||PTX3 interacts with inter-alpha-trypsin inhibitor - Implications for hyaluronan organization and cumulus oophorus expansion|
|Autori:||Scarchilli, L; Camaioni, A; Bottazzi, B; Negri, V; Doni, A; Deban, L; Bastone, A; Salvatori, G; Mantovani, A; Siracusa, G; Salustri, A|
|Appare nelle tipologie:||01 - Articolo su rivista|