请教蛋白质NMR,谢谢

younghe

本人在武汉,急需要做两个蛋白的NMR,有很多问题需要请教各位

做蛋白NMR需要多大的蛋白浓度, 对蛋白的纯度要求是多少?

我只知道有H谱,N谱和C谱,我只是需要做蛋白的空间结构

H谱是不是把蛋白冻干之后加入重水,再测量? 如果做N和C的,是不是要求蛋白中含有13C和15N,这是不是就需要在表达的培养基上加入13C和15N,或者还是直接向溶液里面加入含13C,15N的小分子化合物?

另外,哪个地方可以做蛋白质的NMR,并且对外开放?我想把样品寄过去让别人代做,一般价格是多少?

谢谢!~

我的联系方式:sindamy@yahoo.com.cn

huzhenxin

Several comments:

(1)  If you need protein structure, you can try X-ray crystallography, which will be faster than NMR

(2) If you want to do protein NMR, you need consider how big your protein, if your protein is larger than 25 K, it will be very challenge

(3) protein NMR normally need  at least 0.5 mM protein, the higher, the better, normal requirement is 1 mM, 500 ul

(4) You need label your protein with 2H, which use minimal medium add D2O instead of H2O as solvent, which is for D label

      for N15 label, use minimal medium, use N(15)H4HCl as the only nitrogen source

      for C13 label, use minimal medium, use Glucose (C-13) as the only carbon source

(5) It will take couple weeks to collect all the data, need at least 6 month for an expert to solve the structure. If your protein is very important or very interesting, you

     can try collaborate with Beijing University, they have a Bruker 800MHz. But it will take a lot of time to prepare the sample and much longer time to solve the

     structure.

    Good Luck!