Dear All, I acquired a 3d tocsy-hsqc edited on Bruker machine of small protein in micelles with a mixing time of 70ms. The problem is that I do not observe almoust spin system of each aminoacid, in detail I see only cross peaks between NH-HA. I obtained very good results by 3d Noesy-hsqc edited. How can I do to improve the magnetization transfer along the spin system and see all cross peaks? Do you think that this problem is due to the nature? Thank you in advance\ |
Hi Mario, Relaxation is probably doing you in - the small protein is in a 30+ kD detergent micelle (size varies with detergent, and somewhat with solution conditions), so T2's are too short for a long enough mixing time to get full spin system cross-peaks. I'd suggest backing off to a 40-48 msec mixing time to (hopefully) at least get the H-betas. - Mark |