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. 1999 Jun 15;340(Pt 3):693–701.

N-terminal domains of fibrillin 1 and fibrillin 2 direct the formation of homodimers: a possible first step in microfibril assembly.

T M Trask 1, T M Ritty 1, T Broekelmann 1, C Tisdale 1, R P Mecham 1
PMCID: PMC1220300  PMID: 10359653

Abstract

Aggregation of fibrillin molecules via disulphide bonds is postulated to be an early step in microfibril assembly. By expressing fragments of fibrillin 1 and fibrillin 2 in a mammalian expression system, we found that the N-terminal region of each protein directs the formation of homodimers and that disulphide bonds stabilize this interaction. A large fragment of fibrillin 1 containing much of the region downstream from the N-terminus remained as a monomer when expressed in the same cell system, indicating that this region of the protein lacks dimerization domains. This finding also confirms that the overexpression of fibrillin fragments does not in itself lead to spurious dimer formation. Pulse-chase analysis demonstrated that dimer formation occurred intracellularly, suggesting that the process of fibrillin aggregation is initiated early after biosynthesis of the molecules. These findings also implicate the N-terminal region of fibrillin 1 and fibrillin 2 in directing the formation of a dimer intermediate that aggregates to form the functional microfibril.

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Selected References

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  1. Cleary E. G., Gibson M. A. Elastin-associated microfibrils and microfibrillar proteins. Int Rev Connect Tissue Res. 1983;10:97–209. doi: 10.1016/b978-0-12-363710-9.50009-5. [DOI] [PubMed] [Google Scholar]
  2. Corson G. M., Chalberg S. C., Dietz H. C., Charbonneau N. L., Sakai L. Y. Fibrillin binds calcium and is coded by cDNAs that reveal a multidomain structure and alternatively spliced exons at the 5' end. Genomics. 1993 Aug;17(2):476–484. doi: 10.1006/geno.1993.1350. [DOI] [PubMed] [Google Scholar]
  3. Crouch E., Chang D., Rust K., Persson A., Heuser J. Recombinant pulmonary surfactant protein D. Post-translational modification and molecular assembly. J Biol Chem. 1994 Jun 3;269(22):15808–15813. [PubMed] [Google Scholar]
  4. Del Sal G., Manfioletti G., Schneider C. The CTAB-DNA precipitation method: a common mini-scale preparation of template DNA from phagemids, phages or plasmids suitable for sequencing. Biotechniques. 1989 May;7(5):514–520. [PubMed] [Google Scholar]
  5. Dietz H. C., Cutting G. R., Pyeritz R. E., Maslen C. L., Sakai L. Y., Corson G. M., Puffenberger E. G., Hamosh A., Nanthakumar E. J., Curristin S. M. Marfan syndrome caused by a recurrent de novo missense mutation in the fibrillin gene. Nature. 1991 Jul 25;352(6333):337–339. doi: 10.1038/352337a0. [DOI] [PubMed] [Google Scholar]
  6. Downing A. K., Knott V., Werner J. M., Cardy C. M., Campbell I. D., Handford P. A. Solution structure of a pair of calcium-binding epidermal growth factor-like domains: implications for the Marfan syndrome and other genetic disorders. Cell. 1996 May 17;85(4):597–605. doi: 10.1016/s0092-8674(00)81259-3. [DOI] [PubMed] [Google Scholar]
  7. Kammerer R. A., Schulthess T., Landwehr R., Lustig A., Fischer D., Engel J. Tenascin-C hexabrachion assembly is a sequential two-step process initiated by coiled-coil alpha-helices. J Biol Chem. 1998 Apr 24;273(17):10602–10608. doi: 10.1074/jbc.273.17.10602. [DOI] [PubMed] [Google Scholar]
  8. Kanzaki T., Olofsson A., Morén A., Wernstedt C., Hellman U., Miyazono K., Claesson-Welsh L., Heldin C. H. TGF-beta 1 binding protein: a component of the large latent complex of TGF-beta 1 with multiple repeat sequences. Cell. 1990 Jun 15;61(6):1051–1061. doi: 10.1016/0092-8674(90)90069-q. [DOI] [PubMed] [Google Scholar]
  9. Kielty C. M., Shuttleworth C. A. Synthesis and assembly of fibrillin by fibroblasts and smooth muscle cells. J Cell Sci. 1993 Sep;106(Pt 1):167–173. doi: 10.1242/jcs.106.1.167. [DOI] [PubMed] [Google Scholar]
  10. Lee B., Godfrey M., Vitale E., Hori H., Mattei M. G., Sarfarazi M., Tsipouras P., Ramirez F., Hollister D. W. Linkage of Marfan syndrome and a phenotypically related disorder to two different fibrillin genes. Nature. 1991 Jul 25;352(6333):330–334. doi: 10.1038/352330a0. [DOI] [PubMed] [Google Scholar]
  11. McConnell C. J., DeMont M. E., Wright G. M. Microfibrils provide non-linear elastic behaviour in the abdominal artery of the lobster Homarus americanus. J Physiol. 1997 Mar 1;499(Pt 2):513–526. doi: 10.1113/jphysiol.1997.sp021945. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. McGrath J. A., Sakai L. Y., Eady R. A. Fibrillin immunoreactivity is associated with normal or fragmented elastic microfibrils at the dermal-epidermal junction in recessive dystrophic epidermolysis bullosa. Br J Dermatol. 1994 Oct;131(4):465–471. doi: 10.1111/j.1365-2133.1994.tb08545.x. [DOI] [PubMed] [Google Scholar]
  13. Milewicz D. M., Grossfield J., Cao S. N., Kielty C., Covitz W., Jewett T. A mutation in FBN1 disrupts profibrillin processing and results in isolated skeletal features of the Marfan syndrome. J Clin Invest. 1995 May;95(5):2373–2378. doi: 10.1172/JCI117930. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Pereira L., D'Alessio M., Ramirez F., Lynch J. R., Sykes B., Pangilinan T., Bonadio J. Genomic organization of the sequence coding for fibrillin, the defective gene product in Marfan syndrome. Hum Mol Genet. 1993 Jul;2(7):961–968. doi: 10.1093/hmg/2.7.961. [DOI] [PubMed] [Google Scholar]
  15. Qian R. Q., Glanville R. W. Alignment of fibrillin molecules in elastic microfibrils is defined by transglutaminase-derived cross-links. Biochemistry. 1997 Dec 16;36(50):15841–15847. doi: 10.1021/bi971036f. [DOI] [PubMed] [Google Scholar]
  16. Reinhardt D. P., Keene D. R., Corson G. M., Pöschl E., Bächinger H. P., Gambee J. E., Sakai L. Y. Fibrillin-1: organization in microfibrils and structural properties. J Mol Biol. 1996 Apr 26;258(1):104–116. doi: 10.1006/jmbi.1996.0237. [DOI] [PubMed] [Google Scholar]
  17. Ren Z. X., Brewton R. G., Mayne R. An analysis by rotary shadowing of the structure of the mammalian vitreous humor and zonular apparatus. J Struct Biol. 1991 Feb;106(1):57–63. doi: 10.1016/1047-8477(91)90062-2. [DOI] [PubMed] [Google Scholar]
  18. Ritty T. M., Broekelmann T., Tisdale C., Milewicz D. M., Mecham R. P. Processing of the fibrillin-1 carboxyl-terminal domain. J Biol Chem. 1999 Mar 26;274(13):8933–8940. doi: 10.1074/jbc.274.13.8933. [DOI] [PubMed] [Google Scholar]
  19. Ross R., Bornstein P. The elastic fiber. I. The separation and partial characterization of its macromolecular components. J Cell Biol. 1969 Feb;40(2):366–381. doi: 10.1083/jcb.40.2.366. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Ross R., Fialkow R. J., Altman L. K. The morphogenesis of elastic fibers. Adv Exp Med Biol. 1977;79:7–17. doi: 10.1007/978-1-4684-9093-0_2. [DOI] [PubMed] [Google Scholar]
  21. Saharinen J., Taipale J., Monni O., Keski-Oja J. Identification and characterization of a new latent transforming growth factor-beta-binding protein, LTBP-4. J Biol Chem. 1998 Jul 17;273(29):18459–18469. doi: 10.1074/jbc.273.29.18459. [DOI] [PubMed] [Google Scholar]
  22. Sakai L. Y., Keene D. R., Engvall E. Fibrillin, a new 350-kD glycoprotein, is a component of extracellular microfibrils. J Cell Biol. 1986 Dec;103(6 Pt 1):2499–2509. doi: 10.1083/jcb.103.6.2499. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Sakai L. Y., Keene D. R. Fibrillin: monomers and microfibrils. Methods Enzymol. 1994;245:29–52. doi: 10.1016/0076-6879(94)45004-8. [DOI] [PubMed] [Google Scholar]
  24. Sakai L. Y., Keene D. R., Glanville R. W., Bächinger H. P. Purification and partial characterization of fibrillin, a cysteine-rich structural component of connective tissue microfibrils. J Biol Chem. 1991 Aug 5;266(22):14763–14770. [PubMed] [Google Scholar]
  25. Thurmond F, Trotter J. Morphology and biomechanics of the microfibrillar network of sea cucumber dermis. J Exp Biol. 1996;199(Pt 8):1817–1828. doi: 10.1242/jeb.199.8.1817. [DOI] [PubMed] [Google Scholar]
  26. Tsipouras P., Del Mastro R., Sarfarazi M., Lee B., Vitale E., Child A. H., Godfrey M., Devereux R. B., Hewett D., Steinmann B. Genetic linkage of the Marfan syndrome, ectopia lentis, and congenital contractural arachnodactyly to the fibrillin genes on chromosomes 15 and 5. The International Marfan Syndrome Collaborative Study. N Engl J Med. 1992 Apr 2;326(14):905–909. doi: 10.1056/NEJM199204023261401. [DOI] [PubMed] [Google Scholar]
  27. Wallace R. N., Streeten B. W., Hanna R. B. Rotary shadowing of elastic system microfibrils in the ocular zonule, vitreous, and ligamentum nuchae. Curr Eye Res. 1991 Jan;10(1):99–109. doi: 10.3109/02713689109007614. [DOI] [PubMed] [Google Scholar]
  28. Wright D. W., Mayne R. Vitreous humor of chicken contains two fibrillar systems: an analysis of their structure. J Ultrastruct Mol Struct Res. 1988 Sep;100(3):224–234. doi: 10.1016/0889-1605(88)90039-0. [DOI] [PubMed] [Google Scholar]
  29. Zhang H., Apfelroth S. D., Hu W., Davis E. C., Sanguineti C., Bonadio J., Mecham R. P., Ramirez F. Structure and expression of fibrillin-2, a novel microfibrillar component preferentially located in elastic matrices. J Cell Biol. 1994 Mar;124(5):855–863. doi: 10.1083/jcb.124.5.855. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Zhang H., Hu W., Ramirez F. Developmental expression of fibrillin genes suggests heterogeneity of extracellular microfibrils. J Cell Biol. 1995 May;129(4):1165–1176. doi: 10.1083/jcb.129.4.1165. [DOI] [PMC free article] [PubMed] [Google Scholar]

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