Aycock R S, Raghow R, Stricklin G P, Seyer J M, Kang A H
J Biol Chem. 1986 Oct 25;261(30):14355-60.
We evaluated the effects of a synthetic copy of a highly conserved portion (residues 225-246) of the COOH-propeptide of human pro-alpha 2(I) procollagen on collagen, fibronectin, and total protein synthesis by human fibroblasts. Incubation of COOH-propeptide 225-246 with fibroblasts resulted in a concentration-dependent inhibition of both type I procollagen and fibronectin when compared with controls; a 50% inhibition of both fibronectin and type I collagen was observed at a concentration of 45 microM. Since the overall cellular protein synthesis was only minimally affected, COOH-propeptide appeared to specifically inhibit collagen and fibronectin synthesis. The peptide was nontoxic to cells and the inhibition was completely reversible upon removal of the peptide. We measured the steady-state levels of mRNAs coding for procollagen, fibronectin, and beta-actin by hybridization to specific recombinant cDNA probes; there was no significant change in the steady-state level of mRNAs of the three proteins. These results strongly suggest that the biosynthesis of procollagen and fibronectin in COOH-propeptide-treated cells is inhibited at a post-transcriptional level. These data establish a link between collagen and fibronectin synthesis and further define the important interaction of these molecules in the formation of the extracellular matrix.
我们评估了人原α2(I)型前胶原COOH端前肽高度保守部分(第225 - 246位氨基酸残基)的合成拷贝对人成纤维细胞胶原蛋白、纤连蛋白和总蛋白合成的影响。与对照组相比,将COOH端前肽225 - 246与成纤维细胞一起孵育导致I型前胶原和纤连蛋白均出现浓度依赖性抑制;在45微摩尔浓度时,观察到纤连蛋白和I型胶原均有50%的抑制。由于整体细胞蛋白合成仅受到最小程度的影响,COOH端前肽似乎特异性抑制胶原蛋白和纤连蛋白的合成。该肽对细胞无毒,且去除肽后抑制作用完全可逆。我们通过与特异性重组cDNA探针杂交来测量编码前胶原、纤连蛋白和β - 肌动蛋白的mRNA的稳态水平;这三种蛋白质的mRNA稳态水平没有显著变化。这些结果强烈表明,在COOH端前肽处理的细胞中,前胶原和纤连蛋白的生物合成在转录后水平受到抑制。这些数据建立了胶原蛋白和纤连蛋白合成之间的联系,并进一步明确了这些分子在细胞外基质形成中的重要相互作用。