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CCP4BB  January 2011

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Subject:

Re: NMR structure of 30KD protein, doable?

From:

"Patel, Pryank" <[log in to unmask]>

Reply-To:

Patel, Pryank

Date:

Fri, 21 Jan 2011 20:32:34 +0000

Content-Type:

text/plain

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text/plain (40 lines)

Yes it is, absolutely, but i would say it's a size that "may" require deuteration or other selective labelling methods to obtain the interproton distance restraints required to calculate a structure. The only way to know for sure is to collect some data.

I think even solving a X-ray structure in minutes only happens for the very best cases. For NMR structures it's the same - depends on the quality of the data you have and the person doing it. The software for automatic noesy assignment and structure calculation is relatively robust, and you dont have to worry about growing crystals, but you need to have samples that are stable enough that you can collect the data required (or have multiple samples for all the different experiments).

Say 2-3 weeks for data collection depending on the magnet, another 2-3 weeks for assignment, repeated rounds of structure calculation... A month or two is not out of the question but it's hard to say. 

If you need to deuterate, then you will need to prepare other labelled samples  to get the proton-proton distances for structure calculation. Methyl-labelling is a good start, and there are other experiments that will give you extra data in this case.

CCPN Analysis is one example of software for spectral viewing and assignment. Also have a look at the Aria package for automatic assignment of noesy data and structure calculation. It's basically a frontend for CNS. Many people use Cyana for structure calculation but you need to purchase a licence while Aria and CNS is free.

I'll try and send you more info offlist when I am at a computer.

Best wishes,
Pryank


Sent from my iPhone

On 21 Jan 2011, at 19:56, "Jerry McCully" <[log in to unmask]> wrote:

> Dear ALL;
> 
>        Firstly I am sorry for the offtopic question because it is about NMR.
> 
>        However, I believe that I can get good comments here because all of you are also excellent structural biologists. 
> 
>        During the struggling to crystallize a 30KD protein, I am planning to pursue its NMR structure at the same time. 
> 
>       IS it doable? If so, how long will it take to get the job done? 
> 
>       Thanks to the sophisticated crystallographic software from CCP4, crystallographers can really solve the structure in minutes. 
> 
>       How about NMR structure determination and refinement software? 
> 
>       Welcome any comments and Have a nice weekend.
> 
>   Jerry McCully
> 
> 

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