Evidence for pretranslational regulation of collagen synthesis by procollagen propeptides.
Evidence for pretranslational regulation of collagen synthesis by procollagen propeptides.
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前胶原前肽对胶原合成的翻译前调节的证据。
DOI：
10.1016/s0021-9258(18)67408-3
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发表时间：
1986
期刊：
影响因子：
--
通讯作者：
G. Wu
中科院分区：
文献类型：
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作者：
C. Wu;C. Donovan;G. Wu
在这里，我们提出了前胶原蛋白前肽在胶原蛋白合成的调节中的翻译前作用的证据。氨基和羧基末端的I型前胶原前肽的分离和纯化，从鸡颅盖和肌腱文化。将人肺成纤维细胞（IMR-90）在含有不同浓度的前肽的培养基中孵育。与对照组相比，10 nM的氨基前肽导致胶原蛋白合成减少80%。较高浓度的氨基前肽没有进一步降低胶原蛋白的合成，在整个浓度范围内没有发现对非胶原蛋白合成的显著影响。羧基前肽也抑制胶原合成。在10 nM时，胶原蛋白合成减少30%，40 nM的浓度导致80%的减少。然而，在后一种浓度下，非胶原蛋白合成也受到影响，相对于对照降低了20%。为了评估前肽可能的翻译前作用，用不同浓度的每种前肽处理IMR-90成纤维细胞，并通过与32 P-α 2（I）cDNA探针的斑点杂交测定I型前胶原mRNA的水平。两种前肽均引起I型前胶原mRNA水平的显著浓度依赖性降低。在10 nM时，氨基前肽导致胶原mRNA水平降低55%，而在40 nM时，这些水平与对照相比降低72%。羧基前肽也具有抑制作用，10 nM时mRNA水平降低33%，40 nM时降低73%。信使RNA水平的代表性非胶原蛋白，β-肌动蛋白，不受任何前肽在整个浓度范围。
We present, here, evidence for a pretranslational role of procollagen propeptides in the regulation of collagen synthesis. Amino- and carboxyl-terminal type I procollagen propeptides were isolated and purified from chick calvaria and tendon cultures. Human lung fibroblasts (IMR-90) were incubated in medium containing varying concentrations of propeptides. Amino-propeptides at 10 nM caused an 80% decrease in collagen synthesis compared to control. Higher concentrations of amino-propeptides did not decrease collagen synthesis further and no significant effect on non-collagen synthesis was found throughout the entire concentration range. Carboxyl-propeptides also inhibited collagen synthesis. At 10 nM, collagen synthesis was decreased by 30% and a concentration of 40 nM caused an 80% reduction. However, at the latter concentration non-collagen synthesis was also affected, decreasing by 20% relative to control. To assess possible pretranslational effects of propeptides, IMR-90 fibroblasts were treated with varying concentrations of each propeptide and levels of type I procollagen mRNA was determined by dot hybridization with a 32P-alpha 2(I) cDNA probe. Both propeptides caused significant concentration-dependent decreases in procollagen type I mRNA levels. At 10 nM, the amino-propeptide resulted in a 55% decrease in collagen mRNA levels while at 40 nM these levels decreased by 72% compared to control. Carboxyl-propeptides were also inhibitory, decreasing mRNA levels by 33% at 10 nM and 73% at 40 nM. Messenger RNA levels of a representative noncollagenous protein, beta-actin, were unaffected by either propeptide throughout the concentration range.