Myara I
Laboratoire de Biochimie, Université Paris-Sud, France.
Clin Chim Acta. 1987 Dec;170(2-3):263-70. doi: 10.1016/0009-8981(87)90136-7.
The effect of prolonged preincubation for 24 h at 37 degrees C in the presence of 1 mmol/l manganese at pH 7.8 was investigated on the two forms of human erythrocyte prolidase after their separation by DEAE-Sephadex chromatography. Prolidase I activity, which was eluted in chromatographic fractions of low ionic strength of about 160 mmol/l NaCl, increased after preincubation and this activation was maintained throughout the subsequent steps of chromatofocusing, chromatography on Sephacryl S-200 in tandem with Blue-Sepharose, and Phenyl-Sepharose chromatography. It disappeared after the last step consisting of a second Blue-Sepharose chromatography. When the protein environment was restored, isolated prolidase I was reactivated by preincubation, and as a result, gly-pro procollagen dipeptide became the best substrate. Gly-pro-hyp procollagen tripeptide was also observed to have a strong activator effect. The activity of prolidase II, which was eluted in high ionic strength fractions of about 260 mmol/l NaCl during DEAE-Sephadex chromatography, diminished markedly after preincubation and was very low against the gly-pro substrate. These results seem to indicate that prolidase I is much more active than prolidase II in the intracellular degradation of procollagen.
在pH 7.8条件下，于1 mmol/l锰存在的情况下，在37摄氏度下延长预孵育24小时对通过DEAE - 葡聚糖凝胶色谱分离后的两种形式的人红细胞脯氨酰二肽酶的影响进行了研究。在约160 mmol/l NaCl的低离子强度色谱级分中洗脱的脯氨酰二肽酶I活性，在预孵育后增加，并且在随后的色谱聚焦、与蓝色葡聚糖凝胶串联的Sephacryl S - 200色谱以及苯基葡聚糖凝胶色谱步骤中这种激活作用一直保持。在由第二次蓝色葡聚糖凝胶色谱组成的最后一步之后，它消失了。当恢复蛋白质环境时，分离的脯氨酰二肽酶I通过预孵育重新激活，结果，甘氨酰 - 脯氨酰原胶原二肽成为最佳底物。还观察到甘氨酰 - 脯氨酰 - 羟脯氨酰原胶原三肽具有很强的激活作用。在DEAE - 葡聚糖凝胶色谱期间在约260 mmol/l NaCl的高离子强度级分中洗脱的脯氨酰二肽酶II的活性，在预孵育后明显降低，并且对甘氨酰 - 脯氨酰底物的活性非常低。这些结果似乎表明，在原胶原的细胞内降解中，脯氨酰二肽酶I比脯氨酰二肽酶II活性高得多。
