set up aromatics experiment in VNMRJ2.2

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Dear Sir,

I am new to set up of aromatic assignment experiment. Can you please specify what parameters in VNMRJ 2.2 are important or to be modified for setting up the both aromatic assignment experiments (HB)CB(CGCD)HD and (HB)CB(CGCDCE)HE . What should be the optimum value of constant time in delays and offsets parameters. Whats the optimum value of number of scans and t1 increments

Regards

Arun

aromatics

asked Apr 04 '12 at 00:02

Arun Gupta
11

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Hi Arun,

We ran a (HB)CB(CGCD)HD recently. We used the Biopack version, with a CT of 0.0044 sec. The dof should be 35ppm, and you can set the 13C offset for aromatics to about 125 ppm (Biopack should do this for you). For our protein, 30 increments spanning 3800 Hz was enough.

Note that this is a weak experiment; even with 0.5 mM protein we acquired 2048 transients/increment for a total time of ~36 h.

Hope that helps, Sameer

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answered Apr 05 '12 at 13:00

Sameer Wahid
11

Thanks for your useful suggestion. Should I consider that (HB)CB(CGCDCE)HE would have same sensitivity and require similar experimental time. Did u also run only Aromatic C13-HSQC . Can u suggest how to reference these both spectrum with respect to C13Edit aromatic noesy. As I don't know which amino acid cross peak which will be unique to unambiguously assign in all the 3 spectrum.

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answered Apr 05 '12 at 22:42

Arun Gupta
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Asked: Apr 04 '12 at 00:02

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Last updated: Apr 05 '12 at 22:42

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