title: Deposition of apatite in mineralizing vertebrate extracellular matrices: A model of possible nucleation sites on type I collagen.
abstract: The positions of charged residues in the primary sequence of amino acids comprising the molecular model of type I collagen, the major extracellular protein found in vertebrate tissues, have been earlier characterized by Chapman and Hardcastle [Chapman, J.A., and Hardcastle, R.A. (1974). The staining pattern of collagen fibrils. II. A comparison with patterns computer-generated from the amino acid sequence. Connect. Tissue Res. 2:151-159]. When the sequence of residues is packed in the quarter-staggered arrangement described originally by Hodge and Petruska [Hodge, A.J., and Petruska, J.A. (1963). Recent studies with the electron microscope on ordered aggregates of the tropocollagen macromolecule. In Aspects of Protein Structure, G.N. Ramachandran (ed.) pp. 289-300. New York: Academic Press] in two dimensions and in the quasi-hexagonal model of microfibrillar assembly and molecular packing structure in three dimensions detailed recently by Orgel et al. (Orgel, J.P.R.O., Miller, A., Irving, T.C., Fischetti, R.F., Hammersley, A.P., and Wess, T.J. (2001). The in situ supermolecular structure of type I collagen. Structure 9:1061-1069; Orgel, J.P.R.O., Irving, T.C., Miller, A., and Wess, T.J. (2006). Microfibrillar structure of type I collagen in situ. Proc. Natl. Acad. Sci. U.S.A. 103: 9001-9005], the common sites of charged amino acids, specifically glutamic and aspartic acid, lysine and arginine, and hydroxylysine and histidine, of type I collagen have been examined in the present study and their locations determined in relation to one another. The respective positions of these amino acid residues are notable in several features in two dimensions within a single collagen triple helix as well as in adjacent helices. There are, first, numerous sites in which the same amino acid is adjacent in each of the three collagen helices. Second, many sites exist in which two of the same amino acids and one of the same charge are adjacent in the three helices. Third, the same two or three glutamic and/or aspartic amino acids are found in close proximity to amino acids with their counterparts, aspartic and glutamic acid, respectively. Fourth, several sites occur in which the same two or three amino acids of one charge are present in close proximity to the same two or three amino acids of opposite charge (glutamic acid and lysine or arginine residues or aspartic acid and lysine or arginine residues). Fifth, there are several sites where hydroxylysine contributes charged groups in place of one of the three lysine or arginine residues common in adjacent collagen helices. The strikingly repetitive and close nature of these specific charged groups in two dimensions is even more apparent when the molecular packing structure is investigated in three dimensions. In this instance, the most recent model of Orgel et al. [Orgel, J.P.R.O., Irving, T.C., Miller, A., and Wess, T.J. (2006). Microfibrillar structure of type I collagen in situ. Proc. Natl. Acad. Sci. U.S.A. 103: 9001-9005] has been correlated for the first time with the model of Landis et al. [Landis, W.J., Song, M.J., Leith, A., McEwen, L., and McEwen, B. (1993). Mineral and organic matrix interaction in normally calcifying tendon visualized in three dimensions by high voltage electron microscopic tomography and graphic image reconstruction. J. Struct. Biol. 110: 39-54] showing channels traversing molecular arrays of collagen. Here, many of the charged amino acid sites correspond to the known type I collagen hole zones defined by Hodge and Petruska [Hodge, A.J., and Petruska, J.A. (1963). Recent studies with the electron microscope on ordered aggregates of the tropocollagen macromolecule. In Aspects of Protein Structure, G.N. Ramachandran (ed.) pp. 289-300. New York: Academic Press]. As such, these residues present the locations highly likely to bind Ca(2+) and [Formula: see text] ions in stereochemical configurations that could serve directly as nucleation centers for the subsequent growth and development of apatite crystals representing initial events in vertebrate mineralization. Based on these results, type I collagen appears to provide a molecular framework for direct formation of apatite without the necessary intervention or mediation of other molecules in extracellular matrices of vertebrate calcifying tissues.
authors.affiliation: Department of Pathology and Laboratory Medicine, UMDNJ-Robert Wood Johnson Medical School, Piscataway, NJ, USA.
authors.forename: Frederick H
authors.initials: FH
authors.lastname: Silver
authors.forename: William J
authors.initials: WJ
authors.lastname: Landis
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D001031:Apatites / Q000378:metabolism*; D002113:Calcification, Physiologic; D024042:Collagen Type I / Q000737:chemistry / Q000378:metabolism*; D003460:Crystallization; D005109:Extracellular Matrix / Q000378:metabolism*; D006801:Humans; D008954:Models, Biological; D008958:Models, Molecular; D008969:Molecular Sequence Data; D014714:Vertebrates / Q000378:metabolism*
chemical_list: D001031:Apatites; D024042:Collagen Type I