Collagen type IV is a network-forming collagen that provides support and anchorage to cells. Its basic structural unit is a 410 nm long and 1.5 nm in diameter triple helix, with natural discontinuities in the triple-helical defining Gly-X-Y sequence. The C-terminal globular domain (NC1) in a collagen IV molecule plays an important role in forming networks, and has recently been reported to be structurally triggered by chloride ions to form hexamers outside the cell. How this hexamer assembles in vitro remains unknown. Here, I aim to use atomic force microscopy (AFM) to investigate the molecular basis of collagen type IV network assembly by studying the effects of different solvent conditions on the stability of the NC1 domain. Studying the dissociation of this hexametric domain can shed light onto how it assembles in solution and under what ionic conditions. The flexibility of the collagen type IV molecule is also investigated by performing statistical analysis of AFM-imaged chains and estimating persistence length, a mechanical property that quantifies the flexibility of a polymer. Here, I investigate the effects of triple helix interruptions on the flexibility of the molecule, by comparing collagen type IV to other fibrillar collagens that are continuously triple-helical. In addition, I determine a position-dependent flexibility profile of the molecule showcasing the effects of over-lapping interruptions, from a α1(IV)]2–α2(IV) mouse collagen type IV, on the persistence length.
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