Original Article

The Supramolecular Organization of Fibrillin-rich Microfibrils

Correspondence to: Clair Baldock, Wellcome Trust Centre for Cell-Matrix Research, Schools of Biological Sciences and Medicine, 2.205 Stopford Building, University of Manchester, Manchester, M13 9PT, UK. Tel:0161 275 5756 Fax:0161 275 5752 E-mail:clair.baldock{at}man.ac.uk.

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-Å 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 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.

Key Words: three-dimensional reconstruction, automated electron tomography, fibrillin microfibrils, molecular alignment, scanning transmission electron microscopy mass mapping

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