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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 ' W6 K9 l3 T, { }+ n8 O% p8 ~ experiments detect magnetization that is transferred from a receptor protein C9 e: d, i1 E) c( E$ w ]& I to a bound ligand. Only bound ligands show STD effects. The experiment may be ; S0 a$ W9 O- z2 k% C3 k combined with virtually any other NMR experiment, and therefore is well suitable 7 W) O+ A9 b+ T% b Y# `7 [ to tackle even very complex problems. In particular, in combination with multidimensional ) e) Z4 e1 F7 A! W) k. _ NMR a full characterization of a bound ligand out of a mixture is straightforward. 2 j5 i. [' Z" A8 s STD NMR is extremely robust and gives maximal effects at protein to ligand ratios ) d/ F9 u1 H$ c1 \ greater than ca. 1:100. It follows that less than 1 nmol of protein is necessary j- _5 n2 O! I' Y; o for screening. With the availability of so called cryo probes it will be possible ' W" L! e- B. R6 I7 r. D to work with hundred pmol amounts of protein. The dissociation constant should & N4 ~3 u, Z: F. X" `! M be in the range between nM and mM. Therefore, STD NMR covers at least two orders 2 y* i7 V- ?2 V7 i9 y- R4 U+ p of magnitudes more for dissociation constants than trNOE experiments. From competitive 7 l: i" e% U5 u, G5 \6 u STD experiments dissociation constants may be derived.

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# R* B' L% u2 \4 w% F) B$ F Schematic % y+ Q+ k4 v p( D0 j display of the STD NMR effect. Saturation of the protein leads to a direct saturation $ q3 c6 x, `- o% a% R. m. C of those parts of ligand(s) in direct contact to the protein. By exchange between 1 Z! _) B% O6 l bound and free state the saturation is transported to solution and detected + ?& u8 a2 A1 D8 h2 w- K" A, l by subtracting a spectrum with saturation from a normal spectrum.
/ p- y9 b( q. p: x STD NMR gives precise information about the binding epitope of the ligand. This 4 b. S$ G5 _/ O3 s( c r! _" b, v- m is very important information for the design of a potent drug. The optimal drug ' K! [$ c4 ?) i5 }6 C' Q is of optimal size and optimal shape. The size is deduced from STD NMR, and k3 w0 } n2 d% W" _" P# B! T' ^ the shape is delivered by trNOE experiments.

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