Some properties of the reactive hydroxylysyl residues in collagen: their possible role in nucleation during fibril formation.
Native or heat-denatured collagens were incubated under controlled conditions of temperature and pH with variable molar ratios of KCNO or 2,4,6-trinitrobenzene sulphonic acid. The results obtained suggest that a small number of free amino groups are available for reaction on the native protein, while all the free amino groups react on the denatured protein. The highly reactive free amino groups in the native protein are hydroxylysine residues and have an abnormally low pK of 8.5 which is conformation dependent; this pK becomes normalized upon denaturation of the protein. The reactive hydroxylysines appear to be located in basic regions that could be the nucleation sites needed for fiber formation in the heat-gelation assay; the modified protein does not form stable fibrils upon heating at 37 degrees C and the few fibers formed are not stabilized after reduction with NaBH4. Our results also suggest that the triple helix in collagen is heterogeneous with respect to the reactivity of free amino groups and that several discrete transition temperatures are observed with two main breaks at 30 degrees C and at 37 degrees C, respectively.
D000818:Animals; D003094:Collagen / Q000032:analysis*; D006901:Hydroxylysine / Q000032:analysis*; D007700:Kinetics; D051381:Rats; D013329:Structure-Activity Relationship; D014302:Trinitrobenzenesulfonic Acid; D014783:Viscosity; D014961:X-Ray Diffraction
D006901:Hydroxylysine; D014302:Trinitrobenzenesulfonic Acid; D003094:Collagen