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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 , V0 V) u. k9 h: ~9 X( a& O$ |" e9 \ experiments detect magnetization that is transferred from a receptor protein 3 s7 e1 \2 ?' c3 w* y4 _! o/ K to a bound ligand. Only bound ligands show STD effects. The experiment may be - g$ m) I1 q) Z! g$ v! f combined with virtually any other NMR experiment, and therefore is well suitable : f7 a( s) {$ Q( p' g& y# _' } to tackle even very complex problems. In particular, in combination with multidimensional 4 t4 c# I2 l2 R3 [1 S NMR a full characterization of a bound ligand out of a mixture is straightforward. & s1 ^% v* ?- x4 z) V) |* j STD NMR is extremely robust and gives maximal effects at protein to ligand ratios * y: c3 \& `8 R! | greater than ca. 1:100. It follows that less than 1 nmol of protein is necessary 5 k6 @. b3 T* d for screening. With the availability of so called cryo probes it will be possible 9 |, b6 g% E! @5 D6 p: n9 { to work with hundred pmol amounts of protein. The dissociation constant should # j8 s7 J; u& {9 Q+ Q be in the range between nM and mM. Therefore, STD NMR covers at least two orders ) ~/ L+ [* q6 W# \6 C of magnitudes more for dissociation constants than trNOE experiments. From competitive 0 d2 J4 K2 U# Z. L& _; ]3 ] STD experiments dissociation constants may be derived.

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4 ]* l; T3 e7 X Schematic 3 E' G0 r. `; D- }2 O display of the STD NMR effect. Saturation of the protein leads to a direct saturation l4 Y$ q+ \8 e! P( l# R8 d of those parts of ligand(s) in direct contact to the protein. By exchange between ) x k8 J4 W* {! U& Y bound and free state the saturation is transported to solution and detected - f4 f7 R( G( F# `2 g7 f by subtracting a spectrum with saturation from a normal spectrum.
5 V0 M& ^8 x3 P5 l+ ]8 f STD NMR gives precise information about the binding epitope of the ligand. This ; S: q8 l* O& Z+ s is very important information for the design of a potent drug. The optimal drug % s- F) w ?3 Q5 F is of optimal size and optimal shape. The size is deduced from STD NMR, and * `2 D4 @7 n8 }6 r. w/ @ the shape is delivered by trNOE experiments.

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