title: In vitro fibrillogenesis of collagen type I in varying ionic and pH conditions.
abstract: Collagen is the most abundant protein in the human body, and has primary roles in the formation of tendons, cartilage and bone, it provides mechanical strength to skin and indeed almost every organ and muscle is associated with a layer of collagen. It is thus a key component of the extracellular matrix. Here we have studied the in vitro fibrillogenesis of acetic acid-soluble collagen type I under physiological and varying non-physiological conditions by TEM from negatively stained specimens. At pH 2.5 the collagen heterotrimer remains soluble at increasing buffer concentrations and in the presence of increasing NaCl concentrations. At pH 4.5 molecular aggregates form at low NaCl concentrations, but at higher NaCl concentrations fibrils with a diffuse ~11 nm banding are formed. At pH 7.0, initial molecular aggregates form at low NaCl concentrations that progressively form characteristic ~67 nm D-banded collagen fibrils at intermediate NaCl concentrations that cluster to form thicker multi-fibril D-banded fibres in higher NaCl concentrations. By contrast, increasing concentrations of sodium phosphate at pH 7.0 leads to the formation of flexuous, unbanded fibrils at higher concentrations from the initial, loosely aggregated form of collagen. At higher pHs, the formation of D-banded fibrils is less efficient, particularly at pH 9.0. Thus at neutral pH, the presence of chloride anions, rather than sodium cations, is required for the production of D-banded collagen fibrils; higher than normal physiological chloride concentrations in the form of NaCl or Tris·HCl at neutral pH, but not phosphate buffer, can also lead to the efficient in vitro formation of D-banded collagen fibrils.
authors.affiliation: Institute for Cell and Molecular Biosciences, University of Newcastle, Newcastle-upon-Tyne NE2 4HH, UK. rharris@uni-mainz.de
authors.forename: J Robin
authors.initials: JR
authors.lastname: Harris
authors.forename: Andrei
authors.initials: A
authors.lastname: Soliakov
authors.forename: Richard J
authors.initials: RJ
authors.lastname: Lewis
mesh_terms: D019342:Acetic Acid / Q000378:metabolism; D000818:Animals; D024042:Collagen Type I / Q000187:drug effects*; D006863:Hydrogen-Ion Concentration; D046911:Macromolecular Substances / Q000378:metabolism*; D046529:Microscopy, Electron, Transmission / Q000379:methods; D009994:Osmolar Concentration; D055503:Protein Multimerization / Q000187:drug effects*; D051381:Rats; D012965:Sodium Chloride / Q000378:metabolism
chemical_list: D024042:Collagen Type I; D046911:Macromolecular Substances; D012965:Sodium Chloride; D019342:Acetic Acid