2'（3'）-O-[N-[2-[3-[5-荧光素基]硫脲基]乙基]氨基甲酰基]腺苷5'-三磷酸及其Cr（H₂O）₄和Co（NH₃）₄络合物衍生物是用于标记Na⁺/K⁺-ATP酶ATP结合位点的新型荧光工具。
2'(3')-O-[N- [2- [3- [5-fluoresceinyl] thioureido] ethyl] carbamoyl] adenosine 5'-triphosphate and its Cr(H2O)4 and Co(NH3)4 complex derivatives are new fluorescent tools for labelling ATP binding sites of Na+/K+-ATPase.
作者信息
Linnertz H, Lastres Becker I, Krumscheid R, Amler E, Thoenges D, Schoner W
机构信息
Institute of Biochemistry and Endocrinology, Justus-Liebig-University Giessen, Germany.
出版信息
Physiol Res. 1997;46(5):345-9.
2'(3')-O-[N- [2- [3- [5-fluoresceinyl] thioureido] ethyl] carbamoyl] adenosine 5'-triphosphate (FEDA-ATP), a spectroscopic tool used for studying skeletal muscle myosin ATPase subfragment 1, was applied to Na+/K+-ATPase (EC 3.6.1.37). In contrast to the myosin subfragment, we found that FEDA-ATP is not a substrate for Na+/K+-ATPase. On the other hand, FEDA-ATP showed an affinity for both the low (E2, Kd=200 microM) and the high (E1, Kd=22 microM) affinity ATP-binding sites. When the microscopic affinities of FEDA-ATP were used for calculating the macroscopic affinity in the overall reaction according to Ki=(KdE1*KdE2)1/2, the experimentally measured inhibition constant of 66 microM was obtained. To evoke irreversible binding inhibitors, FEDA-ATP was transferred in its chromium(III) and cobalt(III) complex analogs, which are suitable tools for labelling the ATP binding sites of Na+/K+-ATPase in a specific way.
2'(3')-O-[N-[2-[3-[5-荧光素基]硫脲基]乙基]氨基甲酰基]腺苷5'-三磷酸（FEDA-ATP）是一种用于研究骨骼肌肌球蛋白ATP酶亚片段1的光谱工具，现被应用于Na+/K+-ATP酶（EC 3.6.1.37）。与肌球蛋白亚片段不同，我们发现FEDA-ATP不是Na+/K+-ATP酶的底物。另一方面，FEDA-ATP对低亲和力（E2，Kd = 200 microM）和高亲和力（E1，Kd = 22 microM）的ATP结合位点均表现出亲和力。当根据Ki = (KdE1*KdE2)1/2使用FEDA-ATP的微观亲和力来计算总反应中的宏观亲和力时，得到的实验测量抑制常数为66 microM。为了引发不可逆结合抑制剂，FEDA-ATP被转化为其铬（III）和钴（III）络合物类似物，它们是以特定方式标记Na+/K+-ATP酶ATP结合位点的合适工具。
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