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[求助]NMR检测小分子配体与蛋白相互作用

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发表于 2010-10-18 18:42:26 | 显示全部楼层 |阅读模式

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,

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请教各位大侠,是做小分子的H谱呢?还是做大分子的H谱?

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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?

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各位大侠帮帮忙,不胜感激啊

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发表于 2010-10-18 23:59:29 | 显示全部楼层
应该是用std 实验吧。
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 楼主| 发表于 2010-10-19 12:22:57 | 显示全部楼层
什么事STD实验啊?大侠能详细解释一下吗?小弟初次接触NMR
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发表于 2010-10-19 20:45:31 | 显示全部楼层
同问。蛋白的核磁咋做?
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发表于 2010-10-19 23:54:37 | 显示全部楼层
 

STD NMR

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STD NMR * R+ }& [* ? b+ @* W$ z, q- g, M9 c experiments detect magnetization that is transferred from a receptor protein m: D9 F8 p3 h, ^7 { to a bound ligand. Only bound ligands show STD effects. The experiment may be 9 R# J* @9 q$ E& X. Q5 X combined with virtually any other NMR experiment, and therefore is well suitable - }3 b" R$ S4 O, u+ | m6 |% ^ to tackle even very complex problems. In particular, in combination with multidimensional # }( h* U# p; e" Q+ O& C3 {$ D6 q NMR a full characterization of a bound ligand out of a mixture is straightforward. ) z: c' t1 ^- C/ R STD NMR is extremely robust and gives maximal effects at protein to ligand ratios # U4 e1 p' ^' ?, c8 x5 W- Z7 y greater than ca. 1:100. It follows that less than 1 nmol of protein is necessary q: a; f& Q7 P% `+ w) T for screening. With the availability of so called cryo probes it will be possible 8 z6 S+ B- v, N! u; q to work with hundred pmol amounts of protein. The dissociation constant should : n4 b. q* q4 F0 W0 l# Y9 r be in the range between nM and mM. Therefore, STD NMR covers at least two orders ; L6 V( g/ h) g) {8 X of magnitudes more for dissociation constants than trNOE experiments. From competitive ! F. F' N5 [' [! I' e STD experiments dissociation constants may be derived.

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' E3 b- r: ^- c( G8 t Schematic # @# E2 U; e. C" M! U# l display of the STD NMR effect. Saturation of the protein leads to a direct saturation 1 n" P- m" O; M$ t1 p6 l/ H of those parts of ligand(s) in direct contact to the protein. By exchange between " w# @9 J2 X! x4 \3 O: K; | bound and free state the saturation is transported to solution and detected 4 F a- A7 ?/ v5 z: r6 L by subtracting a spectrum with saturation from a normal spectrum.
/ f* K, Z8 r( w5 p5 b STD NMR gives precise information about the binding epitope of the ligand. This ; t! {) r5 ?4 W is very important information for the design of a potent drug. The optimal drug # `( M# p7 C9 G4 h0 g8 d2 m is of optimal size and optimal shape. The size is deduced from STD NMR, and / {$ ^! ~! J+ ?* D" @, l. x% _ the shape is delivered by trNOE experiments.

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