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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 , P. o9 H7 x J: Q: X9 k7 h* I, F experiments detect magnetization that is transferred from a receptor protein 1 n* |7 o& i N" |. J to a bound ligand. Only bound ligands show STD effects. The experiment may be 1 V1 d3 j- A& B8 l) g combined with virtually any other NMR experiment, and therefore is well suitable 2 H+ l2 A2 P' L9 e% F8 \0 j to tackle even very complex problems. In particular, in combination with multidimensional 0 m1 B3 ^; C" x0 c NMR a full characterization of a bound ligand out of a mixture is straightforward. ' v- Y1 j/ b# C5 |# Z4 ? STD NMR is extremely robust and gives maximal effects at protein to ligand ratios 1 t) E8 @% F/ e$ c7 h) [ greater than ca. 1:100. It follows that less than 1 nmol of protein is necessary 3 s) A& N3 L( K: @, b; Y9 M for screening. With the availability of so called cryo probes it will be possible 4 p2 M# j, d0 w to work with hundred pmol amounts of protein. The dissociation constant should 6 C" @# W0 i4 t8 U' X/ J be in the range between nM and mM. Therefore, STD NMR covers at least two orders + S; R. O9 W: E- [- c0 Z; J of magnitudes more for dissociation constants than trNOE experiments. From competitive 0 r; g0 `# |8 }. m' _ STD experiments dissociation constants may be derived.

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$ M$ T* s; i) p8 J Schematic 4 ^0 g. \& Y2 K. d display of the STD NMR effect. Saturation of the protein leads to a direct saturation 0 a7 Z7 q7 E: \2 H% B of those parts of ligand(s) in direct contact to the protein. By exchange between , p1 {; `2 y2 f3 a7 ^ bound and free state the saturation is transported to solution and detected 2 |4 }8 q/ n+ ~8 \1 C4 b$ _+ { by subtracting a spectrum with saturation from a normal spectrum.
+ O! q0 B. M8 ` STD NMR gives precise information about the binding epitope of the ligand. This $ b& ~2 H/ s& g1 Z# {+ z is very important information for the design of a potent drug. The optimal drug $ b1 `2 V, \! t% P5 f# }6 P. d# N3 M is of optimal size and optimal shape. The size is deduced from STD NMR, and 4 P% [9 [! e8 J the shape is delivered by trNOE experiments.

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