title: Coacervation is promoted by molecular interactions between the PF2 segment of fibrillin-1 and the domain 4 region of tropoelastin.
abstract: In forming elastic fibers, microfibrils act as the scaffold sites for depositing the elastin precursor tropoelastin. We examined key binding interactions that promote massive tropoelastin association through coacervation. Using a segment of the microfibril protein fibrillin-1, PF2, known to bind full-length tropoelastin, we mapped its interaction site to the N-terminal region of tropoelastin bounded by domains 2 and 18. Precise contact residues between domain 4 of tropoelastin and domain 16 of fibrillin-1 were discovered through a novel combination of transglutaminase cross-linking and mass spectroscopy, with contact sites at residues K38 of tropoelastin and Q669 of fibrillin-1. This is the first report of a role for this region of tropoelastin in microfibril interactions. The addition of PF2 thermodynamically facilitated the coacervation of tropoelastin, resulting in smaller changes in entropy and enthalpy values for the coacervating system. A novel multicomponent in vitro tropoelastin assembly reaction system demonstrated that amassed tropoelastin was spatially and preferentially directed to surfaces coated with PF2 as expected for organized three-dimensional distribution during tissue elastogenesis. This study underscores the role of this part of fibrillin-1 as an anchor point for tropoelastin at the microfibril-elastin junction during the initial stages of elastic fiber assembly.
authors.affiliation: School of Molecular and Microbial Biosciences G08, University of Sydney, Sydney, New South Wales, 2006, Australia.
authors.forename: Adam W
authors.initials: AW
authors.lastname: Clarke
authors.forename: Steven G
authors.initials: SG
authors.lastname: Wise
authors.forename: Stuart A
authors.initials: SA
authors.lastname: Cain
authors.forename: Cay M
authors.initials: CM
authors.lastname: Kielty
authors.forename: Anthony S
authors.initials: AS
authors.lastname: Weiss
mesh_terms: D000595:Amino Acid Sequence; D004547:Elastic Tissue / Q000737:chemistry / Q000378:metabolism; D000071838:Fibrillin-1; D000071837:Fibrillins; D006801:Humans; D013058:Mass Spectrometry; D008840:Microfilament Proteins / Q000737:chemistry* / Q000378:metabolism*; D008969:Molecular Sequence Data; D010446:Peptide Fragments / Q000737:chemistry*; D011485:Protein Binding; D011499:Protein Processing, Post-Translational; D017434:Protein Structure, Tertiary; D012996:Solutions; D020349:Surface Plasmon Resonance; D013499:Surface Properties; D013816:Thermodynamics; D017186:Titrimetry; D014333:Tropoelastin / Q000737:chemistry* / Q000378:metabolism*; D054392:Adipokines
chemical_list: D000071838:Fibrillin-1; D000071837:Fibrillins; D008840:Microfilament Proteins; D010446:Peptide Fragments; D012996:Solutions; D014333:Tropoelastin; C000605722:FBN1 protein, human; D054392:Adipokines