Rapanovich I I, Vuĭtsik N R, Tsybanova L Ia
Biokhimiia. 1976 Apr;41(4):740-8.
The specific elution of rabbit skeletal muscle phosphofructokinase (PFK) from DEAE-cellulose is studied in the linear gradient of different allosteric ligands. Citrate and fructose-6-phosphate elute PFK at concentrations of 1.0 and 2.5 mM respectively, i.e. without increasing the ionic strength of the starting buffer (similar to 0.12). The specificity of elution is confirmed by comparison of the ionic strength of these solutions with that of buffer eluting PFK in buffer gradient (mu=0.17) as well as by comparison with the eluting ability of other ligands. Fructose-1,6-diphosphate elutes PFK only at the concentration of 5.5 mM which corresponds to the ionic strength 0.17. MgATP and AMP are inefficient as specific eluents whereas ATP and ADP elute only a small part of PFK with concomitant substantial increase of the ionic strength (up to 0.17--0.18). These results are discussed in terms of a charge compensation mechanism as a result of the displacement of PFK conformers equilibrium under the influence of the allosteric ligands.
研究了在不同变构配体的线性梯度中，兔骨骼肌磷酸果糖激酶（PFK）从DEAE-纤维素上的特异性洗脱情况。柠檬酸和6-磷酸果糖分别在1.0 mM和2.5 mM的浓度下洗脱PFK，即在不增加起始缓冲液离子强度（约为0.12）的情况下。通过将这些溶液的离子强度与在缓冲液梯度中洗脱PFK的缓冲液（μ = 0.17）的离子强度进行比较，以及与其他配体的洗脱能力进行比较，证实了洗脱的特异性。1,6-二磷酸果糖仅在5.5 mM的浓度下洗脱PFK，该浓度对应离子强度0.17。MgATP和AMP作为特异性洗脱剂效率不高，而ATP和ADP仅能洗脱一小部分PFK，同时离子强度大幅增加（高达0.17 - 0.18）。根据变构配体影响下PFK构象体平衡的位移所导致的电荷补偿机制，对这些结果进行了讨论。
