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. 1996 Sep;5(9):1934–1938. doi: 10.1002/pro.5560050922

Conformational flexibility and crystallization of tandemly linked type III modules of human fibronectin.

A Lombardo 1, Y Wang 1, C Z Ni 1, X Dai 1, C D Dickinson 1, R Kodandapani 1, S Chiang 1, C A White 1, F Pio 1, N H Xuong 1, R C Hamlin 1, E Ruoslahti 1, K R Ely 1
PMCID: PMC2143538  PMID: 8880920

Abstract

Fibronectin is a large cell adhesion molecule that is composed of several functional domains. The cell-binding domain that binds to cell surface integrins consists of repeated homologous type III modules. In this study, recombinant fragments from the cell-binding domain of human fibronectin that participate in a newly characterized fibronectin-fibronectin interaction with FNIII1 were crystallized. In each case, the crystals had more than one fibronectin fragment in the asymmetric unit. Crystals of FNIII10-11 grew in the space group C2 with a = 117.1 A, b = 38.6 A, c = 80.6 A, beta = 97.2 degrees, and two molecules in the asymmetric unit. These crystals diffracted to 2.5 A resolution. Fragment FNIII8-11 and a shorter fragment, FNIII8-10, crystallized in hexagonal space groups with large unit cells and two to four molecules per asymmetric unit. Even very large crystals of these fragments did not diffract beyond 4 A. The crystal packing for this collection of fibronectin fragments suggests conformational flexibility between linked type III modules. The functional relevance of this flexibility for elongated versus compact models of the cell-binding domain of fibronectin is discussed.

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Selected References

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  1. Aguirre K. M., McCormick R. J., Schwarzbauer J. E. Fibronectin self-association is mediated by complementary sites within the amino-terminal one-third of the molecule. J Biol Chem. 1994 Nov 11;269(45):27863–27868. [PubMed] [Google Scholar]
  2. Bowditch R. D., Hariharan M., Tominna E. F., Smith J. W., Yamada K. M., Getzoff E. D., Ginsberg M. H. Identification of a novel integrin binding site in fibronectin. Differential utilization by beta 3 integrins. J Biol Chem. 1994 Apr 8;269(14):10856–10863. [PubMed] [Google Scholar]
  3. Dickinson C. D., Veerapandian B., Dai X. P., Hamlin R. C., Xuong N. H., Ruoslahti E., Ely K. R. Crystal structure of the tenth type III cell adhesion module of human fibronectin. J Mol Biol. 1994 Mar 4;236(4):1079–1092. doi: 10.1016/0022-2836(94)90013-2. [DOI] [PubMed] [Google Scholar]
  4. Ely K. R., Kunicki T. J., Kodandapani R. Common molecular scaffold for two unrelated RGD molecules. Protein Eng. 1995 Aug;8(8):823–827. doi: 10.1093/protein/8.8.823. [DOI] [PubMed] [Google Scholar]
  5. Engel J., Odermatt E., Engel A., Madri J. A., Furthmayr H., Rohde H., Timpl R. Shapes, domain organizations and flexibility of laminin and fibronectin, two multifunctional proteins of the extracellular matrix. J Mol Biol. 1981 Jul 25;150(1):97–120. doi: 10.1016/0022-2836(81)90326-0. [DOI] [PubMed] [Google Scholar]
  6. Erickson H. P., Carrell N., McDonagh J. Fibronectin molecule visualized in electron microscopy: a long, thin, flexible strand. J Cell Biol. 1981 Dec;91(3 Pt 1):673–678. doi: 10.1083/jcb.91.3.673. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Ferré-D'Amaré A. R., Burley S. K. Use of dynamic light scattering to assess crystallizability of macromolecules and macromolecular assemblies. Structure. 1994 May 15;2(5):357–359. doi: 10.1016/s0969-2126(00)00037-x. [DOI] [PubMed] [Google Scholar]
  8. Guddat L. W., Herron J. N., Edmundson A. B. Three-dimensional structure of a human immunoglobulin with a hinge deletion. Proc Natl Acad Sci U S A. 1993 May 1;90(9):4271–4275. doi: 10.1073/pnas.90.9.4271. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Huber A. H., Wang Y. M., Bieber A. J., Bjorkman P. J. Crystal structure of tandem type III fibronectin domains from Drosophila neuroglian at 2.0 A. Neuron. 1994 Apr;12(4):717–731. doi: 10.1016/0896-6273(94)90326-3. [DOI] [PubMed] [Google Scholar]
  10. Jones E. Y., Davis S. J., Williams A. F., Harlos K., Stuart D. I. Crystal structure at 2.8 A resolution of a soluble form of the cell adhesion molecule CD2. Nature. 1992 Nov 19;360(6401):232–239. doi: 10.1038/360232a0. [DOI] [PubMed] [Google Scholar]
  11. Kornblihtt A. R., Umezawa K., Vibe-Pedersen K., Baralle F. E. Primary structure of human fibronectin: differential splicing may generate at least 10 polypeptides from a single gene. EMBO J. 1985 Jul;4(7):1755–1759. doi: 10.1002/j.1460-2075.1985.tb03847.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Leahy D. J., Aukhil I., Erickson H. P. 2.0 A crystal structure of a four-domain segment of human fibronectin encompassing the RGD loop and synergy region. Cell. 1996 Jan 12;84(1):155–164. doi: 10.1016/s0092-8674(00)81002-8. [DOI] [PubMed] [Google Scholar]
  13. Leahy D. J., Hendrickson W. A., Aukhil I., Erickson H. P. Structure of a fibronectin type III domain from tenascin phased by MAD analysis of the selenomethionyl protein. Science. 1992 Nov 6;258(5084):987–991. doi: 10.1126/science.1279805. [DOI] [PubMed] [Google Scholar]
  14. Litvinovich S. V., Ingham K. C. Interactions between type III domains in the 110 kDa cell-binding fragment of fibronectin. J Mol Biol. 1995 May 5;248(3):611–626. doi: 10.1006/jmbi.1995.0246. [DOI] [PubMed] [Google Scholar]
  15. Main A. L., Harvey T. S., Baron M., Boyd J., Campbell I. D. The three-dimensional structure of the tenth type III module of fibronectin: an insight into RGD-mediated interactions. Cell. 1992 Nov 13;71(4):671–678. doi: 10.1016/0092-8674(92)90600-h. [DOI] [PubMed] [Google Scholar]
  16. McKeown-Longo P. J., Mosher D. F. Interaction of the 70,000-mol-wt amino-terminal fragment of fibronectin with the matrix-assembly receptor of fibroblasts. J Cell Biol. 1985 Feb;100(2):364–374. doi: 10.1083/jcb.100.2.364. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Morla A., Ruoslahti E. A fibronectin self-assembly site involved in fibronectin matrix assembly: reconstruction in a synthetic peptide. J Cell Biol. 1992 Jul;118(2):421–429. doi: 10.1083/jcb.118.2.421. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Morla A., Zhang Z., Ruoslahti E. Superfibronectin is a functionally distinct form of fibronectin. Nature. 1994 Jan 13;367(6459):193–196. doi: 10.1038/367193a0. [DOI] [PubMed] [Google Scholar]
  19. Mosher D. F., Sottile J., Wu C., McDonald J. A. Assembly of extracellular matrix. Curr Opin Cell Biol. 1992 Oct;4(5):810–818. doi: 10.1016/0955-0674(92)90104-k. [DOI] [PubMed] [Google Scholar]
  20. Obara M., Kang M. S., Yamada K. M. Site-directed mutagenesis of the cell-binding domain of human fibronectin: separable, synergistic sites mediate adhesive function. Cell. 1988 May 20;53(4):649–657. doi: 10.1016/0092-8674(88)90580-6. [DOI] [PubMed] [Google Scholar]
  21. Patthy L. Homology of a domain of the growth hormone/prolactin receptor family with type III modules of fibronectin. Cell. 1990 Apr 6;61(1):13–14. doi: 10.1016/0092-8674(90)90208-v. [DOI] [PubMed] [Google Scholar]
  22. Quade B. J., McDonald J. A. Fibronectin's amino-terminal matrix assembly site is located within the 29-kDa amino-terminal domain containing five type I repeats. J Biol Chem. 1988 Dec 25;263(36):19602–19609. [PubMed] [Google Scholar]
  23. Ruoslahti E. Fibronectin and its receptors. Annu Rev Biochem. 1988;57:375–413. doi: 10.1146/annurev.bi.57.070188.002111. [DOI] [PubMed] [Google Scholar]
  24. Schwarzbauer J. E. Identification of the fibronectin sequences required for assembly of a fibrillar matrix. J Cell Biol. 1991 Jun;113(6):1463–1473. doi: 10.1083/jcb.113.6.1463. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Vrielink A., Beamer L., Le T., Eisenberg D. Crystallization of the chaperone protein SecB. Protein Sci. 1995 Aug;4(8):1651–1653. doi: 10.1002/pro.5560040824. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. de Vos A. M., Ultsch M., Kossiakoff A. A. Human growth hormone and extracellular domain of its receptor: crystal structure of the complex. Science. 1992 Jan 17;255(5042):306–312. doi: 10.1126/science.1549776. [DOI] [PubMed] [Google Scholar]

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