Herold M, Leistler B
Hewlett-Packard GmbH, Waldbronn Analytical Division, Germany.
FEBS Lett. 1992 Aug 10;308(1):26-9. doi: 10.1016/0014-5793(92)81042-k.
Equilibrium dissociation and unfolding of dimeric aspartate aminotransferase from Escherichia coli proceeds via two compact monomeric intermediates which have similar hydrodynamic volumes but different fluorescence properties. We probed binding of the coenzyme pyridoxal 5'-phosphate to these intermediates by coupling fluorescence detection to size-exclusion HPLC. This procedure gave additionally an internal conformational probe of the unfolding transitions of the enzyme. It was shown that the first intermediate, M, is able to bind the coenzyme, whereas the second intermediate, M*, is not. It is likely that M is the correctly folded monomer of the protein.
来自大肠杆菌的二聚天冬氨酸转氨酶的平衡解离和去折叠过程经由两个紧密的单体中间体，它们具有相似的流体力学体积但荧光特性不同。我们通过将荧光检测与尺寸排阻高效液相色谱联用，探测辅酶磷酸吡哆醛与这些中间体的结合。该方法还提供了酶去折叠转变的内部构象探针。结果表明，第一个中间体M能够结合辅酶，而第二个中间体M*则不能。M很可能是该蛋白质正确折叠的单体。
