Fibulin's organization into the extracellular matrix of fetal lung fibroblasts is dependent on fibronectin matrix assembly.
Fibulin is a newly described extracellular matrix (ECM) glycoprotein whose function has not been elucidated. We have observed that cultured fetal lung fibroblasts produce fibulin and have postulated that its expression may be important during lung development. To begin to understand the potential function of fibulin in lung development, we examined its expression and distribution in cultured fetal lung fibroblasts. Immunofluorescence staining of cultured fibroblasts revealed that fibulin was distributed upon their surface in a fibrillar array resembling fibronectin (FN), another ECM glycoprotein expressed by fetal lung fibroblasts and implicated in lung and heart development. Detection of fibulin by immunofluorescence staining of nonpermeabilized cells, its immunoprecipitation from 125I-cell surface-labeled fibroblasts, pulse-chase analysis, and temperature-induced phase separation studies revealed that fibulin is an ECM peripheral membrane protein that is synthesized and secreted by cultured fetal lung fibroblasts shortly after plating and incorporated into their matrix in a divalent cation-dependent manner. Because fibulin co-distributes with both FN and the FN receptor, the integrin alpha 5 beta 1, we examined the possibility that fibulin was interacting with both components. Dissociation of FN receptors from FN fibers with anti-FN receptor antibodies did not affect fibulin's distribution, suggesting that fibulin binds FN and that this interaction is not affected by the state of FN receptor binding. Finally, inhibition of FN matrix assembly prevented the deposition of fibulin, providing further support for FN-fibulin interactions and suggesting that fibulin deposition is dependent on FN matrix assembly.
Roman
J
J
Department of Medicine (111), Atlanta Veterans Affairs Medical Center, Emory University School of Medicine, GA 30033.
McDonald
J A
JA
D002135:Calcium-Binding Proteins / Q000378:metabolism*; D002460:Cell Line; D002462:Cell Membrane / Q000378:metabolism; D005109:Extracellular Matrix / Q000378:metabolism* / Q000648:ultrastructure; D005347:Fibroblasts / Q000166:cytology / Q000648:ultrastructure; D005353:Fibronectins / Q000378:metabolism*; D006801:Humans; D008168:Lung / Q000196:embryology / Q000378:metabolism* / Q000648:ultrastructure; D011233:Precipitin Tests; D013696:Temperature
D002135:Calcium-Binding Proteins; D005353:Fibronectins; C060820:fibulin