Distinction between two molecular species of type V collagen from human post-burn granulation tissues.

Human type V collagen was purified from post-burn granulation tissues, and was demonstrated to exist in two different molecular assemblies consisting of [alpha 1(V)]2 alpha 2(V) and alpha 1(V)alpha 2(V)alpha 3(V) heterotrimers which are designated as type V(112) and V(123) collagens, respectively, in this paper. The two molecular species were separated by salt fractionation at neutral pH under non-denaturing conditions. When crude type V collagen was dialyzed against phosphate-buffered saline at 4 degrees C, mainly collagen V(112) precipitated, leaving collagen V(123) in the solution. Type V(112) collagen, but not type V(123), precipitated at 0.15 M NaCl in 50 mM Tris-HCl buffer (pH 7.5), whereas the type V(123) molecule precipitated at 4.5 M NaCl in the same buffer. When the crude type V collagen was electrophoresed under non-denaturing conditions, two bands were observed; and it was confirmed that the fast-migrating band was composed of [alpha 1(V)]2 alpha 2(V) and the slow-migrating band was alpha 1(V)alpha 2(V)alpha 3(V). Both alpha 1 and alpha 2 chains of V(112) showed biochemical properties that were very similar, if not identical, to those of the corresponding alpha chains of V(123) judging from amino acid compositions, peptide mapping patterns obtained following treatment with cyanogen bromide and lysyl endopeptidase, and periodic acid Schiff and concanavalin A stainings. Alpha 3 chain, in contrast, was distinct from both alpha 1 and alpha 2 chains. The amino acid composition and peptide maps of alpha 3 chain were similar to some extent, but not identical, to those of the alpha 1 chain. The intensity of carbohydrate stainings of the alpha 3 chain was clearly different from that of the alpha 1 chain. The negatively stained segment-long-spacing crystallites of the two molecular species exhibited an identical banding pattern. The crystallite derived from collagen V(112) was usually in a dimeric form exhibiting the C-C terminal junction, but that of collagen V(123) was mostly in a monomeric form. Differences between the two molecular species is ascribed to the presence of the alpha 3 chain in collagen V(123).