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Clin Biochem. 2009 May;42(7-8):713-21. doi: 10.1016/j.clinbiochem.2008.12.020. Epub 2009 Jan 8.

DANCE/fibulin-5 promotes elastic fiber formation in a tropoelastin isoform-dependent manner.

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Department of Clinical Chemistry, Hoshi University School of Pharmacy and Pharmaceutical Sciences, Tokyo, Japan.



To investigate a function of fibulin-5 in the elastic fiber formation, we studied the molecular interactions among elastin, fibrillin-1, and fibulin-5 in the extracellular space and the maturation of tropoelastin using retinal pigment epithelial cells (ARPE-19).


Bacterial recombinant tropoelastin (rTE) was added to ARPE-19 cells overexpressing V5-tagged fibulin-5 (ARPE-Fibulin-5). These elastic fibers were evaluated by immunofluorescence staining, the quantitative analysis of cross-linked amino acids, and semi-quantitative analysis of matrix-associated tropoelastin.


Immunoprecipitation assays revealed that fibulin-5 is able to separately interact with tropoelastin or fibrillin-1 in the culture medium. Moreover, immunofluorescent staining showed that elastin, fibrillin-1, and fibulin-5 co-localize in the extracellular matrix. Desmosine levels were significantly increased in ARPE-Fibulin-5 relative to untransfected cells in spite of equal deposition of tropoelastin by enzyme-linked immunosorbent assay. The addition of a tropoelastin isoform, which lacked the peptide encoded by exon 26A (Delta26A) and could bind to fibulin-5 strongly, led to a larger increase in cross-linking amino acids compared to tropoelastin containing the exon 26A peptide sequence.


These data provide new insights into the initial steps of elastic fiber assembly and identify fibulin-5 and tropoelastin isoforms as potential targets for the regeneration of elastic fibers in vivo.

[Indexed for MEDLINE]

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