Specific Inhibitory Effect of cis-4-Hydroxy-L-Proline on Collagen Synthesis in Growing Mouse Fibroblast 3T6
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概要
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The mouse fibroblast 3T6 cells were incubated in Dulbecco's MEM and the effect of hydroxy-proline analogue, cis-4-hydroxy-L-proline (cis-HYP), was examined on the sequence of collagen formation in the growing cells.<BR>Collagen was labeled with <SUP>14</SUP>C-L-proline (PRO) in the medium and the fate of radioactivity in the collagen was paralelled with that of <I>trans</I>-hydroxyproline (HYP).<BR>Incorporation of <SUP>14</SUP>C-L-PRO to the cellular or medium protein was inhibited by cis-HYP to the extent of 85% by 600 μg/ml of the analogue and intracellular accumulation of 14C-L-PRO-peptide did not increase in the presence of cis-HYP. Non-collagenuos protein synthesis was impaired slightly by high cocentration of cis-HYP. Specificity of cis-HYP on the collagen synthesis further proved by the selective digestion of the labeled protein with purified Clostridium collagenase.<BR>Effect of cis-HYP on the cell growth and the viability was not significant up to the concentra-tion of 600μg/ml.<BR>Labeled intracellular collagen synthesized in the presence of cis-HYP was extruded to the culture medium at the same rate as the normal, and the rate of extrusion of the normal collagen from the cell was not interfered by cis-HYP in the medium.<BR>These result suggested that cis-HYP selectively inhibits de novo biosynthesis of collagen in the 3T6 cells.
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