Effect of streptococcal lipoteichoic acid on prolyl hydroxylase activity as related to collagen formation in mouse fibroblast monolayers.
Dried and wet mouse fibroblast monolayers with labeled collagenous substrate were used to study the effects of lipoteichoic acid (LTA) on cellular prolyl hydroxylase activity. LTA is a scavenger of cations, and Fe2+ is essential for prolyl hydroxylase activity. Surprisingly, addition of LTA to dried monolayers resulted in increased prolyl hydroxylase activity, whereas preincubation of Fe2+ with LTA only negated this increase. However, significant inhibition of enzyme activity by wet monolayers occurred whether LTA was added directly to the test system or whether it was used after preincubation with Fe2+. These data suggest that LTA causes membrane perturbations. Also, that the binding of LTA to the membrane of dried and wet monolayers appears to be decidedly different when based on the subsequent availability of Fe2+ for cellular prolyl hydroxylase activity. The ability of LTA to act as a cationic exchanger and the presence of intracellular Fe2+ inaccessible to LTA probably accounted for the lack of complete inhibition of prolyl hydroxylase activity by this amphiphile in the wet cell system. Considerably less iron was needed to negate the partial inhibition of prolyl hydroxylase activity by LTA in viable cells than was needed to restore the increased enzyme activity by this amphiphile in equivalent dried preparations. These and other results showed that, although LTA does not affect collagen polypeptide chain formation in wet monolayers, its involvement at the molecular level does result in a marked decrease in the hydroxylation of collagenous peptidyl prolyl residues through LTA interaction with Fe2+. This reduction in prolyl hydroxylase activity equaled the reduction in hydroxylation of collagenous protein in fibroblast monolayers caused by LTA reported earlier (O. Leon and C. Panos, Infect. Immun. 40:785-794, 1983). Therefore, these data suggest that partial inhibition of prolyl hydroxylase activity is directly related to the synthesis of defective collagen by wet fibroblast monolayers exposed to minute amounts of group A, type 12 streptococcal LTA. Use of LTA also showed that complete inhibition of hydroxyproline formation is not required for the continued formation and accumulation of defective collagenous protein by these monolayers.
D000818:Animals; D002470:Cell Survival; D002478:Cells, Cultured; D003094:Collagen / Q000096:biosynthesis*; D005296:Ferrous Compounds / Q000494:pharmacology; D005347:Fibroblasts / Q000201:enzymology; D006900:Hydroxylation; D008070:Lipopolysaccharides; D051379:Mice; D010712:Phosphatidic Acids / Q000494:pharmacology*; D011393:Procollagen-Proline Dioxygenase / Q000032:analysis*; D013297:Streptococcus pyogenes / Q000032:analysis*; D013682:Teichoic Acids / Q000494:pharmacology*
D005296:Ferrous Compounds; D008070:Lipopolysaccharides; D010712:Phosphatidic Acids; D013682:Teichoic Acids; C009900:lipoteichoic acid; D003094:Collagen; D011393:Procollagen-Proline Dioxygenase