找回密码
 马上注册

扫一扫,访问微社区

搜索
热搜: mestrenova
查看: 2720|回复: 4

[求助]NMR检测小分子配体与蛋白相互作用

[复制链接]
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,

$ X0 U7 d9 l8 L0 A0 L9 c, _1 F4 G

请教各位大侠,是做小分子的H谱呢?还是做大分子的H谱?

# q: k) D0 l' M% i7 [4 ^' Y

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?

) V( }3 n8 k1 m, \" {# n( o0 z, z3 r

各位大侠帮帮忙,不胜感激啊

回复

使用道具 举报

发表于 2010-10-18 23:59:29 | 显示全部楼层
应该是用std 实验吧。
回复 支持 反对

使用道具 举报

 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
回复 支持 反对

使用道具 举报

发表于 2010-10-19 20:45:31 | 显示全部楼层
同问。蛋白的核磁咋做?
回复 支持 反对

使用道具 举报

发表于 2010-10-19 23:54:37 | 显示全部楼层
 

STD NMR

6 r8 `" m1 E3 m3 Q/ c

STD NMR 8 k ~) f0 T+ ], q: l experiments detect magnetization that is transferred from a receptor protein 5 y" z! H' Q) F% y( I9 r to a bound ligand. Only bound ligands show STD effects. The experiment may be , C0 U# e9 @* Z. p6 Q combined with virtually any other NMR experiment, and therefore is well suitable ' `' ]# b! v' e% I! s& T( ? to tackle even very complex problems. In particular, in combination with multidimensional ' D& c+ P0 v* [( t' C- p: t4 ^ NMR a full characterization of a bound ligand out of a mixture is straightforward. ! m0 e6 [- g0 S STD NMR is extremely robust and gives maximal effects at protein to ligand ratios : e: z% Y7 ]6 d6 T4 b greater than ca. 1:100. It follows that less than 1 nmol of protein is necessary , g! m6 B7 V( f; W9 q6 X for screening. With the availability of so called cryo probes it will be possible 9 C W G& z3 `! U. ?2 v `- A to work with hundred pmol amounts of protein. The dissociation constant should ; w7 T' i8 L9 ]7 h be in the range between nM and mM. Therefore, STD NMR covers at least two orders ( V) v! M, _# {' |4 w1 f of magnitudes more for dissociation constants than trNOE experiments. From competitive - A$ p+ U' [# G+ P0 M- d4 }. n STD experiments dissociation constants may be derived.

) ~0 B% ?' U, ?" V


3 o" z4 s c7 j1 R

, l$ a* L0 J: _& @ m2 c. I% l Schematic * M& f) r, Q# X0 W display of the STD NMR effect. Saturation of the protein leads to a direct saturation " K5 r- E$ r0 l! } of those parts of ligand(s) in direct contact to the protein. By exchange between 7 z: H% B5 A* x. R& c3 K bound and free state the saturation is transported to solution and detected 8 t# h& \: O! t, N. f by subtracting a spectrum with saturation from a normal spectrum.
; ]" G2 d+ S& }& q STD NMR gives precise information about the binding epitope of the ligand. This , J+ s' Z" M' v. o is very important information for the design of a potent drug. The optimal drug + m, \& R4 M7 H' U" K4 Z is of optimal size and optimal shape. The size is deduced from STD NMR, and 5 o4 X; P5 m* c the shape is delivered by trNOE experiments.

回复 支持 反对

使用道具 举报

您需要登录后才可以回帖 登录 | 马上注册

本版积分规则

Archiver|手机版|中国核磁共振论坛

GMT+8, 2025-11-23 20:49

Powered by Discuz! X3.5

© 2001-2024 Discuz! Team.

快速回复 返回顶部 返回列表