title: Decreased synthesis and increased intracellular degradation of newly synthesized collagen in freshly isolated chick tendon cells incubated with monensin.
abstract: The synthesis and secretion of procollagen in embryonic chick tendon fibroblasts in suspension culture were inhibited with the carboxylic ionophore monensin. The synthesis of procollagen was inhibited by 50% in a 2-h exposure to 0.1 microM monensin and was inhibited by 70% in a 6-h exposure to 0.1 microM monensin. Secretion of procollagen was inhibited by greater than 90% in the 0.1 microM monensin-treated cultures and was totally inhibited by higher doses of the reagent. A cellular pool of collagenase-digestible peptides was demonstrated in the control cells, the level of which was elevated 3-4 times in the monensin-treated cultures. In order to determine whether the secretory and synthesis block caused by monensin inhibited intracellular degradation of newly synthesized collagen, the hydroxy[14C]proline in degraded collagen fragments present in control and monensin-treated cultures was determined and compared to the total hydroxy[14C]proline synthesized in each culture. The intracellular degradation of newly synthesized, pulse-labeled collagen was shown to proceed at rates comparable to those seen in the control cultures. The monensin-treated cells degraded pulse-labeled newly synthesized collagen nearly twice as long as the controls, resulting in an overall increase in the fraction of newly synthesized collagen that was degraded. These findings suggest that force generation in the activated cross-bridge cycle may occur as a result of an actin-attached cross-bridge transition between these two orientations.
mesh_terms: D000818:Animals; D002478:Cells, Cultured; D002642:Chick Embryo; D003094:Collagen / Q000096:biosynthesis / Q000378:metabolism*; D007700:Kinetics; D008985:Monensin / Q000494:pharmacology*; D011392:Proline / Q000378:metabolism; D013710:Tendons / Q000187:drug effects / Q000378:metabolism*
chemical_list: D003094:Collagen; D008985:Monensin; D011392:Proline