[求助]NMR检测小分子配体与蛋白相互作用

rushui117

刚接触NMR，想用来检测一下一种三个苯环的小分子配体与一种三万KDa的蛋白的相互作用，

' x5 l$ U4 p* i( o1 r+ w

请教各位大侠，是做小分子的H谱呢？还是做大分子的H谱?

4 ]! k2 q, w$ J7 d6 A3 Y7 E% W

做的话对于溶剂有什么要求吗？蛋白用什么溶剂呢？小分子用DMSO溶解可以吗？

0 Z" Q/ \; w/ H+ g" b

各位大侠帮帮忙，不胜感激啊

syflu

应该是用std 实验吧。

rushui117

什么事STD实验啊？大侠能详细解释一下吗？小弟初次接触NMR

考洛希

同问。蛋白的核磁咋做？

syflu

 

STD NMR

STD NMR & a* k, t: u7 X0 J experiments detect magnetization that is transferred from a receptor protein to a bound ligand. Only bound ligands show STD effects. The experiment may be + u) H. q2 l; R+ r; L combined with virtually any other NMR experiment, and therefore is well suitable 9 H- s5 G' @ p V, N1 e% m' M" w* K to tackle even very complex problems. In particular, in combination with multidimensional 9 c; V) P. Q( \2 k& G NMR a full characterization of a bound ligand out of a mixture is straightforward. 0 d! x& u7 G( ?, |; r$ N STD NMR is extremely robust and gives maximal effects at protein to ligand ratios greater than ca. 1:100. It follows that less than 1 nmol of protein is necessary : o9 G3 i2 C/ D7 {4 V# h. x6 V for screening. With the availability of so called cryo probes it will be possible 4 ~+ R, E- h! c; C% P/ \- R! T to work with hundred pmol amounts of protein. The dissociation constant should be in the range between nM and mM. Therefore, STD NMR covers at least two orders $ i1 q) L* a) d$ Y' S# y of magnitudes more for dissociation constants than trNOE experiments. From competitive 8 P7 {) H5 c/ Q% o- M1 s/ Z STD experiments dissociation constants may be derived.

i; a4 ~1 M, A7 A+ E: p0 d
5 E- ^4 g% X0 P6 t J, u8 O' m Schematic display of the STD NMR effect. Saturation of the protein leads to a direct saturation # b, i9 n: V7 L; l+ f4 Q of those parts of ligand(s) in direct contact to the protein. By exchange between 4 N. b; M# g7 \ bound and free state the saturation is transported to solution and detected + q2 k$ K1 k) ?9 W" Q by subtracting a spectrum with saturation from a normal spectrum.
$ f9 }/ ^7 U4 A0 k9 ] STD NMR gives precise information about the binding epitope of the ligand. This is very important information for the design of a potent drug. The optimal drug , e8 |4 a* r+ P1 u: q is of optimal size and optimal shape. The size is deduced from STD NMR, and the shape is delivered by trNOE experiments.