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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 6 A+ A2 M* j$ ]7 m% B3 G experiments detect magnetization that is transferred from a receptor protein 5 r1 H8 {" y/ s! P' F' o( A to a bound ligand. Only bound ligands show STD effects. The experiment may be 2 a5 S. L' `9 z; H* E combined with virtually any other NMR experiment, and therefore is well suitable ' d" \) Y/ O- | to tackle even very complex problems. In particular, in combination with multidimensional 5 P+ T5 ~' S. v" `% L3 @ NMR a full characterization of a bound ligand out of a mixture is straightforward. " u# Z {% H: R! \5 q STD NMR is extremely robust and gives maximal effects at protein to ligand ratios * f9 a1 ?6 {$ l9 [0 n4 ~ greater than ca. 1:100. It follows that less than 1 nmol of protein is necessary , |- S+ j v N for screening. With the availability of so called cryo probes it will be possible ; d9 L1 O* ]. S% d to work with hundred pmol amounts of protein. The dissociation constant should - }" }% C% I/ q2 Q7 f8 h be in the range between nM and mM. Therefore, STD NMR covers at least two orders 1 |+ Z" P, }$ {7 q* ?2 y& X8 J of magnitudes more for dissociation constants than trNOE experiments. From competitive / V4 W: i! k- H& H5 n/ P" G STD experiments dissociation constants may be derived.

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* n/ d( F' q+ `6 _% {- [ Schematic 4 V# ]" J- a& O! R0 O* Q display of the STD NMR effect. Saturation of the protein leads to a direct saturation - ~ I: c# ?, K& H1 e of those parts of ligand(s) in direct contact to the protein. By exchange between ) D6 n4 E1 W- j, _ bound and free state the saturation is transported to solution and detected 7 K, A$ s' f# }1 u, Y: X by subtracting a spectrum with saturation from a normal spectrum.
/ B5 J; E2 b( l3 v STD NMR gives precise information about the binding epitope of the ligand. This , v5 [1 }" X' c: M' A is very important information for the design of a potent drug. The optimal drug ! I5 A! c% h4 s. c' J Q, m is of optimal size and optimal shape. The size is deduced from STD NMR, and " v, |3 F; |, W) G, ]9 } the shape is delivered by trNOE experiments.

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