Early fibrillin-1 assembly monitored through a modifiable recombinant cell approach

Dirk Hubmacher, Eric Bergeron, Christine Fagotto-Kaufmann, Lynn Sakai, Dieter P. Reinhardt

Research output: Contribution to journalArticle

9 Citations (Scopus)

Abstract

Fibrillin proteins constitute the backbone of extra-cellular macromolecular microfibrils. Mutations in fibrillins cause heritable connective tissue disorders, including Marfan syndrome, dominant Weill-Marchesani syndrome, and stiff skin syndrome. Fibronectin provides a critical scaffold for microfibril assembly in cell culture models. Full length recombinant fibrillin-1 was expressed by HEK 293 cells, which deposited the secreted protein in a punctate pattern on the cell surface. Cocultured fibroblasts consistently triggered assembly of recombinant fibrillin-1, which was dependent on a fibronectin network formed by the fibroblasts. Deposition of recombinant fibrillin-1 on fibronectin fibers occurred first in discrete packages that subsequently extended along fibronectin fibers. Mutant fibrillin-1 harboring either a cysteine 204 to serine mutation or a RGD to RGA mutation which prevents integrin binding, did not affect fibrillin-1 assembly. In conclusion, we developed a modifiable recombinant full-length fibrillin-1 assembly system that allows for rapid analysis of critical roles in fibrillin assembly and functionality. This system can be used to study the contributions of specific residues, domains, or regions of fibrillin-1 to the biogenesis and functionality of microfibrils. It provides also a method to evaluate disease-causing mutations, and to produce microfibril-containing matrices for tissue engineering applications, for example, in designing novel vascular grafts or stents.

Original languageEnglish (US)
Pages (from-to)1456-1468
Number of pages13
JournalBiomacromolecules
Volume15
Issue number4
DOIs
StatePublished - Apr 14 2014
Externally publishedYes

Fingerprint

Fibronectins
Microfibrils
Fibroblasts
Mutation
Proteins
Weill-Marchesani Syndrome
Stents
Fibers
Scaffolds (biology)
Tissue engineering
Cell culture
Grafts
Integrins
Serine
Marfan Syndrome
Cysteine
Skin
HEK293 Cells
Tissue Engineering
Fibrillin-1

ASJC Scopus subject areas

  • Bioengineering
  • Materials Chemistry
  • Polymers and Plastics
  • Biomaterials
  • Medicine(all)

Cite this

Hubmacher, D., Bergeron, E., Fagotto-Kaufmann, C., Sakai, L., & Reinhardt, D. P. (2014). Early fibrillin-1 assembly monitored through a modifiable recombinant cell approach. Biomacromolecules, 15(4), 1456-1468. https://doi.org/10.1021/bm5000696

Early fibrillin-1 assembly monitored through a modifiable recombinant cell approach. / Hubmacher, Dirk; Bergeron, Eric; Fagotto-Kaufmann, Christine; Sakai, Lynn; Reinhardt, Dieter P.

In: Biomacromolecules, Vol. 15, No. 4, 14.04.2014, p. 1456-1468.

Research output: Contribution to journalArticle

Hubmacher, D, Bergeron, E, Fagotto-Kaufmann, C, Sakai, L & Reinhardt, DP 2014, 'Early fibrillin-1 assembly monitored through a modifiable recombinant cell approach', Biomacromolecules, vol. 15, no. 4, pp. 1456-1468. https://doi.org/10.1021/bm5000696
Hubmacher, Dirk ; Bergeron, Eric ; Fagotto-Kaufmann, Christine ; Sakai, Lynn ; Reinhardt, Dieter P. / Early fibrillin-1 assembly monitored through a modifiable recombinant cell approach. In: Biomacromolecules. 2014 ; Vol. 15, No. 4. pp. 1456-1468.
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