Incorporation of in vitro formed 4D-staggered linear polymers of collagen into D-periodic fibrils.
We have previously demonstrated in vitro formation of acid-labile, unidirectional, and 4D-staggered linear polymers of collagen molecules at low ionic strength, low temperature, and pH 4.5. The possibility was investigated in this paper whether the 4D-staggered linear polymers thus formed could be incorporated into native-type, D-periodic collagen fibrils. For this purpose, acid-labile, 4D-staggered linear polymers of collagen were formed in vitro by dialysing monomeric collagen solution against acetate buffer, pH 4.5, and the 4D-staggered association was chemically fixed with glutaraldehyde. The 4D-staggered linear polymers gave fibrous, or end-overlapped segment-long-spacing (SLS) forms when dialysed against ATP/acetic acid solution. 4D-Periodic SLS forms were dialysed against 0.14 M NaCl-30 mM phosphate buffer (pH 7.4) at 37 degrees C. Samples were taken at intervals and examined under an electron microscope. After 4D-periodic SLS being loosened, D-periodic fine fibril structure appeared within 4 hours of dialysis. After longer incubation, the preference of 4D-periodicity completely disappeared and D-periodic polar fibrils of collagen were formed with negligible fine filaments in the background. It is concluded that in vitro formed 4D-staggered linear polymers can be incorporated into D-periodic collagen fibrils under physiological conditions.
D000818:Animals; D003094:Collagen / Q000378:metabolism*; D006863:Hydrogen-Ion Concentration; D009994:Osmolar Concentration; D011108:Polymers / Q000378:metabolism; D051381:Rats; D013696:Temperature
D011108:Polymers; D003094:Collagen