The supramolecular organization of fibrillin-rich microfibrils.
We propose a new model for the alignment of fibrillin molecules within fibrillin microfibrils. Automated electron tomography was used to generate three-dimensional microfibril reconstructions to 18.6-A resolution, which revealed many new organizational details of untensioned microfibrils, including heart-shaped beads from which two arms emerge, and interbead diameter variation. Antibody epitope mapping of untensioned microfibrils revealed the juxtaposition of epitopes at the COOH terminus and near the proline-rich region, and of two internal epitopes that would be 42-nm apart in unfolded molecules, which infers intramolecular folding. Colloidal gold binds microfibrils in the absence of antibody. Comparison of colloidal gold and antibody binding sites in untensioned microfibrils and those extended in vitro, and immunofluorescence studies of fibrillin deposition in cell layers, indicate conformation changes and intramolecular folding. Mass mapping shows that, in solution, microfibrils with periodicities of <70 and >140 nm are stable, but periodicities of approximately 100 nm are rare. Microfibrils comprise two in-register filaments with a longitudinal symmetry axis, with eight fibrillin molecules in cross section. We present a model of fibrillin alignment that fits all the data and indicates that microfibril extensibility follows conformation-dependent maturation from an initial head-to-tail alignment to a stable approximately one-third staggered arrangement.
Baldock
C
C
Wellcome Trust Centre for Cell-Matrix Research, Schools of Biological Sciences and Medicine, University of Manchester, Manchester, M13 9PT, United Kingdom. clair.baldock@man.ac.uk
Koster
A J
AJ
Ziese
U
U
Rock
M J
MJ
Sherratt
M J
MJ
Kadler
K E
KE
Shuttleworth
C A
CA
Kielty
C M
CM
D000595:Amino Acid Sequence; D000818:Animals; D000906:Antibodies / Q000276:immunology; D001331:Automation; D001666:Binding Sites, Antibody; D001704:Biopolymers / Q000737:chemistry / Q000276:immunology / Q000378:metabolism; D002417:Cattle; D002478:Cells, Cultured; D004815:Epidermal Growth Factor / Q000737:chemistry; D000071837:Fibrillins; D005347:Fibroblasts; D005455:Fluorescent Antibody Technique; D017979:Gold Colloid / Q000378:metabolism; D006801:Humans; D007091:Image Processing, Computer-Assisted; D020894:Microfibrils / Q000737:chemistry* / Q000276:immunology / Q000378:metabolism / Q000648:ultrastructure*; D008840:Microfilament Proteins / Q000737:chemistry / Q000276:immunology / Q000378:metabolism / Q000648:ultrastructure*; D017348:Microscopy, Electron, Scanning Transmission; D008958:Models, Molecular; D008969:Molecular Sequence Data; D009129:Muscle Tonus; D020836:Protein Structure, Quaternary; D017434:Protein Structure, Tertiary; D014054:Tomography / Q000379:methods
D000906:Antibodies; D001704:Biopolymers; D000071837:Fibrillins; D017979:Gold Colloid; D008840:Microfilament Proteins; D004815:Epidermal Growth Factor