Altered rate of fibronectin matrix assembly by deletion of the first type III repeats

Jan L. Sechler, Yoshikazu Takada, Jean E. Schwarzbauer

研究成果: 雜誌貢獻文章

115 引文 (Scopus)

摘要

The assembly of fibronectin (FN) into a fibrillar matrix is a complex stepwise process that involves binding to integrin receptors as well as interactions between FN molecules. To follow the progression of matrix formation and determine the stages during which specific domains function, we have developed cell lines that lack an endogenous FN matrix but will form fibrils when provided with exogenous FN. Recombinant FNs (recFN) containing deletions of either the RGD cell-binding sequence (RGD-) or the first type III repeats (FNΔIII1-7) including the III1 FN binding site were generated with the baculovirus insect cell expression system. After addition to cells, recFN matrix assembly was monitored by indirect immunofluorescence and by insolubility in the detergent deoxycholate (DOC). In the absence of any native FN, FNΔIII1-7 was assembled into fibrils and was converted into DOC-insoluble matrix. This process could be inhibited by the amino- terminal 70 kD fragment of FN, showing that FNΔIII1-7 follows an assembly pathway similar to FN. The progression of FNΔIII1-7 assembly differed from native FN in that the recFN became DOC-insoluble more quickly. In contrast, RGD recFNs were not formed into fibrils except when added in combination with native FN. These results show that the RGD sequence is essential for the initiation step but fibrils can form independently of the III1-7 modules. The altered rate of FNΔIII1-7 assembly suggests that one function of the missing repeats might be to modulate an early stage of matrix formation.

原文英語
頁(從 - 到)573-583
頁數11
期刊Journal of Cell Biology
134
發行號2
DOIs
出版狀態已發佈 - 七月 1996
對外發佈Yes

指紋

Fibronectins
Deoxycholic Acid
Baculoviridae
Indirect Fluorescent Antibody Technique
Integrins
Detergents
Insects
Binding Sites
Cell Line

ASJC Scopus subject areas

  • Cell Biology

引用此文

Altered rate of fibronectin matrix assembly by deletion of the first type III repeats. / Sechler, Jan L.; Takada, Yoshikazu; Schwarzbauer, Jean E.

於: Journal of Cell Biology, 卷 134, 編號 2, 07.1996, p. 573-583.

研究成果: 雜誌貢獻文章

Sechler, Jan L. ; Takada, Yoshikazu ; Schwarzbauer, Jean E. / Altered rate of fibronectin matrix assembly by deletion of the first type III repeats. 於: Journal of Cell Biology. 1996 ; 卷 134, 編號 2. 頁 573-583.
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