IMPROVED CHROMATOGRAPHIC PURIFICATION OF HUMAN AND BOVINE TYPE-V COLLAGEN SUB-MOLECULAR SPECIES AND THEIR SUBUNIT CHAINS FROM CONVENTIONAL CRUDE PREPARATIONS - APPLICATION TO CELL-SUBSTRATUM ADHESION ASSAY FORHUMAN UMBILICAL VEIN ENDOTHELIAL-CELL

Two human type V collagen sub-molecular species, designated [alpha 1(V )](2) alpha 2(V) and alpha 1(V)alpha 2(V)alpha 3(V), were purified chr omatographically from a commercially available preparation, in which c ystine-rich collagenous contaminants were contained, with a column pac ked with Fractogel EMD SO,. From bovine crude preparations, the [alpha 1(V)](2) alpha 2(V) form free from the collagenous contaminants was p urified. Type V collagen subunit chains were isolated from each type V collagen molecule by anion-exchange HPLC with a Bakerbond PEI Scout c olumn. The highly purified human type V collagen molecules and their s ubunit chains were used to examine the inhibitory effect on human umbi lical vein endothelial cell proliferation. It was confirmed that the a lpha 1(V) chain has inhibitory activity and it was found that the inhi bitory effect of the [alpha 1(V)](2) alpha 2(V) form is stronger than that of the alpha 1(V)alpha 2(V)alpha 3(V) form and that the alpha 3(V ) chain has no inhibitory activity.