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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?

( U* p5 t+ a, P- K

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 + q8 |" Q# ?) C ~! v experiments detect magnetization that is transferred from a receptor protein ' i8 A; R% c& w$ ?1 i& l to a bound ligand. Only bound ligands show STD effects. The experiment may be " b* ^6 d, \3 q0 j; \ B; M combined with virtually any other NMR experiment, and therefore is well suitable 9 S1 ?; _8 j6 f$ B# y6 W, l: ~3 h. } to tackle even very complex problems. In particular, in combination with multidimensional : X1 F2 e2 P2 P3 H$ I NMR a full characterization of a bound ligand out of a mixture is straightforward. 2 Z9 r/ Y) a, C0 a STD NMR is extremely robust and gives maximal effects at protein to ligand ratios + w9 I+ |5 g1 F: Q/ ] greater than ca. 1:100. It follows that less than 1 nmol of protein is necessary ; t6 {3 a: ~# T& z: q( g for screening. With the availability of so called cryo probes it will be possible % B! f& h! i) A$ F- w to work with hundred pmol amounts of protein. The dissociation constant should . s" o% X" ]" h0 T8 J/ [ be in the range between nM and mM. Therefore, STD NMR covers at least two orders # R/ h% @8 k2 E: ]+ a of magnitudes more for dissociation constants than trNOE experiments. From competitive 4 ^9 m& A2 o3 ^3 v0 h2 ` STD experiments dissociation constants may be derived.

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/ i8 `# u% L- s" z' p Schematic 9 ~2 ]. B ]$ r! e9 Z" X+ q display of the STD NMR effect. Saturation of the protein leads to a direct saturation % j- |1 W; M* p" v4 r, G of those parts of ligand(s) in direct contact to the protein. By exchange between ' z7 ]. E0 m, x: o2 |6 ?/ [$ E bound and free state the saturation is transported to solution and detected 3 A# c: [" G w/ v by subtracting a spectrum with saturation from a normal spectrum.
2 D6 p+ m5 ^( f$ k6 V: s STD NMR gives precise information about the binding epitope of the ligand. This , H, R. u$ Q$ \( P! f" P is very important information for the design of a potent drug. The optimal drug # T" {, s. I: b$ D1 \: H' F; b' J is of optimal size and optimal shape. The size is deduced from STD NMR, and 0 [; _/ k6 u4 u& t; q5 M, D the shape is delivered by trNOE experiments.

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