Aida T, Asano G
Nihon Ika Daigaku Zasshi. 1989 Aug;56(4):361-4. doi: 10.1272/jnms1923.56.361.
Prolyl hydroxylase (EC 1.14.11.2) synthesis was examined in the cultured corneas of 17-day-old chick embryos which were cultured in F-12 medium supplemented with fetal calf serum. The amount of prolyl hydroxylase in cells was measured by immunoelectrophoresis using antibodies directed against this enzyme. The results indicated that the amounts of prolyl hydroxylase depended on cell density. The cells were harvested at the early-log phase and plated at high cell density to mimic the culture condition of cells at the stationary phase (crowding). The amount of 35S-prolyl hydroxylase indicated a two-fold increase in the rate of prolyl hydroxylase synthesis after crowding. By use of 32P-labeled cDNA for the beta-subunit of prolyl hydroxylase a proportionate increase of the mRNA was noted in accordance with the increase in the rate of prolyl hydroxylase synthesis, suggesting that this enzyme synthesis is regulated at the transcriptional level.
在添加胎牛血清的F-12培养基中培养17日龄鸡胚的角膜，检测脯氨酰羟化酶（EC 1.14.11.2）的合成。使用针对该酶的抗体通过免疫电泳测量细胞中脯氨酰羟化酶的量。结果表明，脯氨酰羟化酶的量取决于细胞密度。在对数早期收获细胞，并以高细胞密度接种，以模拟细胞处于静止期（拥挤）的培养条件。35S-脯氨酰羟化酶的量表明拥挤后脯氨酰羟化酶合成速率增加了两倍。通过使用脯氨酰羟化酶β亚基的32P标记cDNA，发现mRNA随着脯氨酰羟化酶合成速率的增加而相应增加，表明该酶的合成在转录水平受到调控。
