Print

Print


Dear All,
Does anyone know of a facility
that provides amino acid analysis for determination of SeMet
incorporation in a recombinant protein?
We're looking for some advice
about how to proceed with a structure we're working on.  Our
protein is 350 amino acids (10 methionine are present) and naturally binds magnesium.  We
have a SeMet data set at peak that goes down to  5.4 angstroms. 
We tried to find the heavy atom position with Shelxcde program. The
log file is given as below.

Resl.   Inf - 8.0 - 6.0 - 5.6 - 5.4 -
5.2 - 5.0 - 4.8 - 4.6 - 4.4 - 4.2 - 3.98 
N(data)     529   643   241    29    
0     0     0     0     0     0     0 
<I/sig>    63.5  19.5  10.6 
10.0   0.0   0.0   0.0   0.0   0.0   0.0   0.0 
%Complete  94.1  98.9  99.2  18.0  
0.0   0.0   0.0   0.0   0.0   0.0   0.0 
<d"/sig>   3.06  1.24  1.07 1.21 

I am also attaching the .lst and .res file and
from that we were considering that only one selenomethionine
position.We have one monomer per AU
(-unfortunately, MR is not working for this project). The space group
is P3 .  We also have a native set down to 3.4 angstroms. 
While we understand that we may need more phasing information  we're
wondering if anyone might have some other suggestions or insights
about how we can move forward given the data that we currently have. 
Can we extend the phases with our native data set. Thanks in advance
for any advice.

regards

 
Vijay