Domains 16 and 17 of tropoelastin in elastic fibre formation.

Naturally occurring mutations are useful in identifying domains that are important for protein function. We studied a mutation in the elastin gene, 800-3G>C, a common disease allele for SVAS (supravalvular aortic stenosis). We showed in primary skin fibroblasts from two different SVAS families that this mutation causes skipping of exons 16-17 and results in a stable mRNA. Tropoelastin lacking domains 16-17 (Delta16-17) was synthesized efficiently and secreted by transfected retinal pigment epithelium cells, but showed the deficient deposition into the extracellular matrix compared with normal as demonstrated by immunofluorescent staining and desmosine assays. Solid-phase binding assays indicated normal molecular interaction of Delta16-17 with fibrillin-1 and fibulin-5. However, self-association of Delta16-17 was diminished as shown by an elevated coacervation temperature. Moreover, negative staining electron microscopy confirmed that Delta16-17 was deficient in forming fibrillar polymers. Domain 16 has high homology with domain 30, which can form a beta-sheet structure facilitating fibre formation. Taken together, we conclude that domains 16-17 are important for self-association of tropoelastin and elastic fibre formation. This study is the first to discover that domains of elastin play an essential role in elastic fibre formation by facilitating homotypic interactions.

[1]  Fumiaki Sato,et al.  The characteristics of elastic fiber assembled with recombinant tropoelastin isoform. , 2006, Clinical biochemistry.

[2]  J. Cirulis,et al.  Structural determinants of cross-linking and hydrophobic domains for self-assembly of elastin-like polypeptides. , 2005, Biochemistry.

[3]  A. Weiss,et al.  Coacervation is promoted by molecular interactions between the PF2 segment of fibrillin-1 and the domain 4 region of tropoelastin. , 2005, Biochemistry.

[4]  Hiroshi Wachi,et al.  Development of a new in vitro model of elastic fiber assembly in human pigmented epithelial cells. , 2005, Clinical biochemistry.

[5]  A. Shuttleworth,et al.  Fibulin-5 interacts with fibrillin-1 molecules and microfibrils. , 2005, The Biochemical journal.

[6]  Anthony S Weiss,et al.  Molecular Basis of Elastic Fiber Formation , 2004, Journal of Biological Chemistry.

[7]  M. Miao,et al.  Sequence and Structure Determinants for the Self-aggregation of Recombinant Polypeptides Modeled after Human Elastin* , 2003, Journal of Biological Chemistry.

[8]  K. Woodhouse,et al.  Recombinant human elastin polypeptides self‐assemble into biomaterials with elastin‐like properties , 2003, Biopolymers.

[9]  Hiroshi Wachi,et al.  Domains in Tropoelastin That Mediate Elastin Depositionin Vitro and in Vivo * , 2003, The Journal of Biological Chemistry.

[10]  M. Rock,et al.  Microfibril-associated Glycoprotein-2 Interacts with Fibrillin-1 and Fibrillin-2 Suggesting a Role for MAGP-2 in Elastic Fiber Assembly* , 2002, The Journal of Biological Chemistry.

[11]  Paul Coucke,et al.  Homozygosity for a missense mutation in fibulin-5 (FBLN5) results in a severe form of cutis laxa. , 2002, Human molecular genetics.

[12]  B. Meier,et al.  The molecular structure of spider dragline silk: Folding and orientation of the protein backbone , 2002, Proceedings of the National Academy of Sciences of the United States of America.

[13]  D. Chitayat,et al.  Connection between elastin haploinsufficiency and increased cell proliferation in patients with supravalvular aortic stenosis and Williams-Beuren syndrome. , 2002, American journal of human genetics.

[14]  C. Bellingham,et al.  Elastin as a self-organizing biomaterial: use of recombinantly expressed human elastin polypeptides as a model for investigations of structure and self-assembly of elastin. , 2002, Philosophical transactions of the Royal Society of London. Series B, Biological sciences.

[15]  Masashi Yanagisawa,et al.  Fibulin-5 is an elastin-binding protein essential for elastic fibre development in vivo , 2002, Nature.

[16]  Tasuku Honjo,et al.  Fibulin-5/DANCE is essential for elastogenesis in vivo , 2002, Nature.

[17]  A. Weiss,et al.  Hydrophobic Domains of Human Tropoelastin Interact in a Context-dependent Manner* , 2001, The Journal of Biological Chemistry.

[18]  A. Weiss,et al.  Protein Interaction Studies of MAGP-1 with Tropoelastin and Fibrillin-1* , 2001, The Journal of Biological Chemistry.

[19]  J. Belmont,et al.  Supravalvular aortic stenosis: genetic and molecular dissection of a complex mutation in the elastin gene , 2001, Human Genetics.

[20]  D. Milewicz,et al.  Genetic disorders of the elastic fiber system. , 2000, Matrix biology : journal of the International Society for Matrix Biology.

[21]  J. Rosenbloom,et al.  Interaction of Tropoelastin with the Amino-terminal Domains of Fibrillin-1 and Fibrillin-2 Suggests a Role for the Fibrillins in Elastic Fiber Assembly* , 2000, The Journal of Biological Chemistry.

[22]  A. Munnich,et al.  Isolated supravalvular aortic stenosis: functional haploinsufficiency of the elastin gene as a result of nonsense-mediated decay , 2000, Human Genetics.

[23]  A. Weiss,et al.  Deficient coacervation of two forms of human tropoelastin associated with supravalvular aortic stenosis. , 1999, European journal of biochemistry.

[24]  R. Mecham,et al.  Characterization of an in vitro model of elastic fiber assembly. , 1999, Molecular biology of the cell.

[25]  D. Milewicz,et al.  Processing of the Fibrillin-1 Carboxyl-terminal Domain* , 1999, The Journal of Biological Chemistry.

[26]  S. Thibodeau,et al.  Supravalvular aortic stenosis: a splice site mutation within the elastin gene results in reduced expression of two aberrantly spliced transcripts , 1999, Human Genetics.

[27]  R. Mecham,et al.  Novel arterial pathology in mice and humans hemizygous for elastin. , 1998, The Journal of clinical investigation.

[28]  Dean Y. Li,et al.  Elastin is an essential determinant of arterial morphogenesis , 1998, Nature.

[29]  A. Weiss,et al.  Coacervation characteristics of recombinant human tropoelastin. , 1997, European journal of biochemistry.

[30]  D. Keene,et al.  The association of human fibulin-1 with elastic fibers: an immunohistological, ultrastructural, and RNA study. , 1995, The journal of histochemistry and cytochemistry : official journal of the Histochemistry Society.

[31]  P. Robson,et al.  Characterization of lamprin, an unusual matrix protein from lamprey cartilage. Implications for evolution, structure, and assembly of elastin and other fibrillar proteins. , 1993, The Journal of biological chemistry.

[32]  R. Mecham,et al.  The cysteine residues in the carboxy terminal domain of tropoelastin form an intrachain disulfide bond that stabilizes a loop structure and positively charged pocket. , 1992, Biochemical and biophysical research communications.

[33]  J. Rosenbloom,et al.  Cloning of full-length elastin cDNAs from a human skin fibroblast recombinant cDNA library: further elucidation of alternative splicing utilizing exon-specific oligonucleotides. , 1988, The Journal of investigative dermatology.

[34]  L Peltonen,et al.  Alternative splicing of human elastin mRNA indicated by sequence analysis of cloned genomic and complementary DNA. , 1987, Proceedings of the National Academy of Sciences of the United States of America.

[35]  D. Volpin,et al.  Relevance of aggregation properties of tropoelastin to the assembly and structure of elastic fibers. , 1986, Journal of ultrastructure and molecular structure research.

[36]  J. Rosenbloom,et al.  The role of the carboxy terminus of tropoelastin in its assembly into the elastic fiber. , 1999, Connective tissue research.

[37]  R. Mecham,et al.  Desmosine radioimmunoassay as a means of studying elastogenesis in cell culture. , 1981, Connective tissue research.