Krupianko V I
Biokhimiia. 1976 Apr;41(4):703-7.
The effect of pH on the kinetic parameters (Km and Ki) for extracellular acid Penicillium brevicompactum RNAse (pH max 4.7+/-0.1), non-specific to the chemical nature of nucleic bases, was studied. The pKm--pH dependence curve showed bends within the following intervals of pH: 3.5--4.0 and 5.6--6.0 (upward side concavity) and 6.2--6.8 (downward side concavity). The pKi--pH dependence for adenosine-3'-monophosphate as an inhibitor is identical to the pH dependence on pKm for the substrate. On the other hand, the pKi--pH dependence curves obtained for the base-free inhibitors (ribose-5'-monophosphate, or phosphate (adenosine) show no bends within the pH intervals of 3.0--4.0 and 5.6--7.0 respectively. A possibility is discussed of the presence of a carboxylic (pK 3.58+/-0.1) and two imidazole groups (pK 6.42+/-0.1--a weakly protonated and 5.8+/-+/-0.1--a strongly protonated group) in the RNAse active site and their participation in the formation of the RNAse-nucleotide (RNAse-substrate) complex.
研究了pH对短密青霉胞外酸性核糖核酸酶(pH最大值4.7±0.1)动力学参数(Km和Ki)的影响,该酶对核苷酸碱基的化学性质无特异性。pKm-pH依赖性曲线在以下pH区间出现弯曲:3.5-4.0和5.6-6.0(向上凹)以及6.2-6.8(向下凹)。作为抑制剂的3'-单磷酸腺苷的pKi-pH依赖性与底物的pKm-pH依赖性相同。另一方面,对于无碱基抑制剂(5'-单磷酸核糖或磷酸腺苷)获得的pKi-pH依赖性曲线在3.0-4.0和5.6-7.0的pH区间内分别没有弯曲。讨论了核糖核酸酶活性位点中存在一个羧基(pK 3.58±0.1)和两个咪唑基团(pK 6.42±0.1——弱质子化和5.8±0.1——强质子化基团)及其参与核糖核酸酶-核苷酸(核糖核酸酶-底物)复合物形成的可能性。