HMQC and HSQC experiments with water flip-back optimized for large proteins |
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| Authors: | Patrik Andersson Bernard Gsell Beat Wipf Hans Senn Gottfried Otting |
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| Affiliation: | (1) Department of Medical Biochemistry and Biophysics, Karolinska Institute, Doktorsringen 9A, S-171 77 Stockholm, Sweden;(2) Zentrale Forschungseinheit, F. Hoffmann-La Roche AG, CH-4002 Basel, Switzerland |
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| Abstract: | An HMQC experiment is proposed, dubbed FHMQC, where water flip-back is achieved by a single water-selective pulse preceding the basic HMQC pulse sequence. The scheme is demonstrated with a 15N, 1H-HMQC spectrum of uniformly 15N/2H-labelled S. aureus DNA gyrase B with a molecular weight of 45 kDa for the unlabelled protein. The sensitivity of the experiment is improved compared to that of an FHSQC spectrum. It is further shown that the original FHSQC experiment can be shortened by the use of bipolar gradients. Relaxation times of different 15N magnetizations and coherences were measured. The new FHMQC scheme is implemented in 3D NOESY-15N-HMQC and 3D15 N-HMQC-NOESY-15N-HMQC pulse sequences which are demonstrated with a 24 kDa fragment of uniformly 15N/13C/2H-labelled S. aureus DNA gyrase B. |
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| Keywords: | 3D 15N-HMQC-NOESY-15N-HMQC 3D NOESY-15N-HMQC DNA gyrase B deuteration FHMQC FHSQC HMQC HSQC 15N relaxation |
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