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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 3 z/ w. {2 t I4 k8 s2 C experiments detect magnetization that is transferred from a receptor protein ' n. c6 t! v7 G* h9 Q$ ?% h5 X1 A% Q to a bound ligand. Only bound ligands show STD effects. The experiment may be 4 k2 b7 r5 i j2 E9 P" B combined with virtually any other NMR experiment, and therefore is well suitable 2 K+ ?. o0 H$ U% f to tackle even very complex problems. In particular, in combination with multidimensional ) p6 j/ J7 D0 p. B& B) C0 f NMR a full characterization of a bound ligand out of a mixture is straightforward. 2 t8 E9 p, X6 u! X/ _4 R STD NMR is extremely robust and gives maximal effects at protein to ligand ratios 6 L- N! F" \& Y- S greater than ca. 1:100. It follows that less than 1 nmol of protein is necessary ( d; A* I$ R" a- L- F2 D for screening. With the availability of so called cryo probes it will be possible 7 E( P* i) b/ M: B to work with hundred pmol amounts of protein. The dissociation constant should : l0 b2 `$ d3 c, N. b be in the range between nM and mM. Therefore, STD NMR covers at least two orders ; `4 S/ L6 t) o5 j1 b# } of magnitudes more for dissociation constants than trNOE experiments. From competitive # R; |3 o0 `7 j- x- M2 L4 v, ^ STD experiments dissociation constants may be derived.

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/ G/ i5 Z" Z% n5 V Schematic : V/ {+ O8 T2 ]4 s6 E6 e display of the STD NMR effect. Saturation of the protein leads to a direct saturation ; r" R( Q8 Z4 E7 i$ r7 F, N of those parts of ligand(s) in direct contact to the protein. By exchange between 1 K8 C4 W k+ ~% | bound and free state the saturation is transported to solution and detected 7 j: H1 W2 E4 b by subtracting a spectrum with saturation from a normal spectrum.
$ c. H7 Z2 o) z STD NMR gives precise information about the binding epitope of the ligand. This 9 @: G' ] e( G$ A v# H' c" u is very important information for the design of a potent drug. The optimal drug 1 O+ z3 e, Q X; ` W, }" N is of optimal size and optimal shape. The size is deduced from STD NMR, and - N" J ] n' S' {! x/ B the shape is delivered by trNOE experiments.

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