title: Identification of a major microfibril-associated glycoprotein-1-binding domain in fibrillin-2.
abstract: Using yeast two-hybrid, ligand blotting, and solid phase binding assays, we have shown that microfibril-associated glycoprotein-1 (MAGP-1) interacts with the 8-cysteine motif of fibrillin-2 encoded by exon 24. Binding to this sequence was demonstrated for full-length MAGP-1 as well as for the MAGP-1 matrix-binding domain encoded by exons 7 and 8. The matrix-binding domain, but not the full-length protein, also bound to regions of fibrillin-2 defined by exons 16 and 17, exon 20, and exons 23 and 24. Interestingly, no binding was detected to sequences near the N or C terminus where MAGP-1 and MAGP-2, respectively, were shown to interact with fibrillin-1. The localization of MAGP-1 binding to the 8-Cys domain encoded by exon 24 suggests that the bead structure of microfibrils consists of exon 24 and portions of the central region of fibrillin-2. Exon 24 in fibrillin lies in the region of the molecule where mutations produce the most severe phenotypes associated with Marfan syndrome (fibrillin-1) and congenital contractural arachnodactyly (fibrillin-2). It is possible that these mutations alter the ability of fibrillin to bind MAGP-1, which may contribute to the severity of the disease.
authors.affiliation: Department of Cell Biology, Washington University School of Medicine, St. Louis, Missouri 63110, USA.
authors.forename: Claudio C
authors.initials: CC
authors.lastname: Werneck
authors.forename: Barbara Crippes
authors.initials: BC
authors.lastname: Trask
authors.forename: Thomas J
authors.initials: TJ
authors.lastname: Broekelmann
authors.forename: Timothy M
authors.initials: TM
authors.lastname: Trask
authors.forename: Timothy M
authors.initials: TM
authors.lastname: Ritty
authors.forename: Fernando
authors.initials: F
authors.lastname: Segade
authors.forename: Robert P
authors.initials: RP
authors.lastname: Mecham
mesh_terms: D000818:Animals; D001665:Binding Sites / Q000235:genetics; D016466:CHO Cells; D002417:Cattle; D003285:Contractile Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D006224:Cricetinae; D005091:Exons; D016326:Extracellular Matrix Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D000071837:Fibrillins; D008840:Microfilament Proteins / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D011485:Protein Binding; D017434:Protein Structure, Tertiary; D000072260:RNA Splicing Factors
chemical_list: D003285:Contractile Proteins; D016326:Extracellular Matrix Proteins; D000071837:Fibrillins; D008840:Microfilament Proteins; D000072260:RNA Splicing Factors; C016598:microfibrillar protein