Open AccessSynchrotron radiation x-ray scattering in the early stages of in vitro collagen fibril formation.
Author(s) -
Gerardo Suárez,
Arnold L. Oronsky,
J. Bordas,
Michel H. J. Koch
Publication year - 1985
Publication title -
proceedings of the national academy of sciences of the united states of america
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 5.011
H-Index - 771
eISSN - 1091-6490
pISSN - 0027-8424
DOI - 10.1073/pnas.82.14.4693
Subject(s) - fibril , chemistry , scattering , polymerization , phase (matter) , dynamic light scattering , crystallography , synchrotron radiation , small angle x ray scattering , biophysics , in vitro , synchrotron , analytical chemistry (journal) , materials science , chromatography , biochemistry , optics , organic chemistry , nanotechnology , physics , nanoparticle , biology , polymer
The time course of in vitro collagen fibril formation was monitored by synchrotron radiation x-ray scattering. Collagen polymerization was induced by a temperature jump from 4 degrees C to 32 degrees C and the solution scattering pattern was recorded continuously with a time resolution of a few seconds. The scattered intensity increased as soon as the final temperature was attained, without discernible lag phase, when the collagen concentration was about 0.56 mg/ml, whereas turbidimetric measurements revealed a lag phase of ca. 2.5 min at 1.05 mg/ml. A direct correlation was found between the temperature and both the rate of formation and the total amount of early aggregates. The formation of these aggregates was only partially reversed by lowering the temperature, except when the collagen had been prevented from forming Schiff base-mediated intermolecular crosslinks by sodium borohydride reduction. In this case, formation of aggregates was completely reversible. We conclude that the aggregates that appear to correspond to the subfibrils proposed on the basis of independent methods are formed and simultaneously crosslinked in the early phases of in vitro collagen self-assembly.