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 for human 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 chromatographically from a commercially available preparation, in which cystine-rich collagenous contaminants were contained, with a column packed with Fractogel EMD SO3-. From bovine crude preparations, the [alpha 1(V)]2 alpha 2(V) form free from the collagenous contaminants was purified. Type V collagen subunit chains were isolated from each type V collagen molecule by anion-exchange HPLC with a Bakerbond PEI Scout column. The highly purified human type V collagen molecules and their subunit chains were used to examine the inhibitory effect on human umbilical vein endothelial cell proliferation. It was confirmed that the alpha 1(V) chain has inhibitory activity and it was found that the inhibitory 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.
Sato
K
K
Department of Food Science and Nutrition, Kyoto Prefectural University, Japan.
Taira
T
T
Takayama
R
R
Ohtsuki
K
K
Kawabata
M
M
D000818:Animals; D002417:Cattle; D002448:Cell Adhesion / Q000187:drug effects*; D002455:Cell Division / Q000187:drug effects; D002851:Chromatography, High Pressure Liquid / Q000379:methods; D003094:Collagen / Q000737:chemistry / Q000302:isolation & purification* / Q000494:pharmacology; D004591:Electrophoresis, Polyacrylamide Gel; D004730:Endothelium, Vascular / Q000166:cytology*; D005260:Female; D006801:Humans; D046911:Macromolecular Substances; D010920:Placenta / Q000737:chemistry; D013710:Tendons / Q000737:chemistry; D014471:Umbilical Veins
D046911:Macromolecular Substances; D003094:Collagen