Background: Type I collagen contains specific lysine and hydroxylysine residues that are critical in the formation of intermolecular cross-links crucial for the normal configuration and stability of the 67 nm axial repeat of collagen fibrils in the extracellular matrix. The major cross-linkage sites are believed to occur between the non-helical terminal regions (telopeptides) and helical segments of adjacent collagen molecules. In this X-ray fibre diffraction study the tissue has been maintained in the hydrated fibrillar state, whilst detailed structural information was obtained using highly collimated synchrotron radiation.
Results: The axial component of the X-ray diffraction patterns extends more than twice as far in reciprocal space than that of any already published. The structure-factor phases were calculated using the multiple isomorphous addition method, avoiding model-based approaches, and produced an electron-density profile of the molecular arrangement projected on to the fibre axis to 0.54 nm resolution. This corresponds to the phasing of 124 orders of the meridional diffraction pattern.
Conclusions: The axially projected electron-density profile and the electron-density difference maps showed that both the N- and C-terminal telopeptides are contracted structures. This profile puts narrow constraints on the possible conformations of the C-terminal telopeptide; the best fit to the electron-density profile is when the α1 chains adopt a folded conformation with a sharp hairpin turn around residues 13 and 14 of the 25-residue telopeptide. Our results reveal for the first time the location, parallel to the fibril axis, of the intermolecular cross-links in normal hydrated tissue. These cross-links are essential for the biological function of the tissue.
