In:
Cells Tissues Organs, S. Karger AG, Vol. 183, No. 1 ( 2006), p. 41-52
Kurzfassung:
The extracellular matrix surrounding avian oocytes, referred to as the perivitelline membrane (PL), exhibits a three-dimensional network of fibrils between granulosa cells and the oocyte. We previously reported that one of its components, ZPC, is synthesized in granulosa cells that are specifically incorporated into the PL; this incorporation might be mediated by a specific interaction with 〈 i 〉 ZPB1, 〈 /i 〉 another PL constituent, which is synthesized in the liver. In order to extend our previous findings, we established an expression system for quail 〈 i 〉 ZPB1 〈 /i 〉 using a mammalian cell line, and several 〈 i 〉 ZPB1 〈 /i 〉 mutants lacking the zona pellucida (ZP) domain or the glutamine-rich repeat region were produced. Western blot analysis of the immunoprecipitated materials with anti-ZPC antiserum indicated that 〈 i 〉 ZPB1 〈 /i 〉 was coimmunoprecipitated with the antiserum in the presence of ZPC. Ligand blotting also revealed the specific binding of ZPC and 〈 i 〉 ZPB1 〈 /i 〉 and indicated that the binding of these two components might be mediated via an ionic interaction. An analysis using recombinant 〈 i 〉 ZPB1 〈 /i 〉 demonstrated that the 〈 i 〉 ZPB1 〈 /i 〉 lacking the ZP domain did not bind to ZPC, whereas the mutant missing the glutamine-rich repeat region retained its capacity for binding. Furthermore, although the 〈 i 〉 ZPB1 〈 /i 〉 lacking the N-terminal half of the ZP domain was able to bind to ZPC, the deletion of the C-terminal half completely abolished 〈 i 〉 ZPB1 〈 /i 〉 binding to ZPC. These results suggested that the C-terminal half of the ZP domain of 〈 i 〉 ZPB1 〈 /i 〉 contains a binding site for ZPC, and that it appears to be involved in insoluble PL fibril formation in the quail ovary.
Materialart:
Online-Ressource
ISSN:
1422-6405
,
1422-6421
Sprache:
Englisch
Verlag:
S. Karger AG
Publikationsdatum:
2006
ZDB Id:
1481840-1
SSG:
12
