Application of multiple-quantum line narrowing with simultaneous 1H and 13C constant-time scalar-coupling evolution in PFG-HACANH and PFG-HACA(CO)NH triple-resonance experiments

Authors: Swapna G.V.T.; Rios C.B.; Shang Z.; Montelione G.T.

Source: Journal of Biomolecular NMR, Volume 9, Number 1, January 1997 , pp. 105-111(7)

Publisher: Springer

Abstract:

Many triple-resonance experiments make use of one-bond heteronuclear scalar couplings to establish connectivities among backbone and/or side-chain nuclei. In medium-sized (15–30 kDa) proteins, short transverse relaxation times of C single-quantum states limit signal-to-noise (S/N) ratios. These relaxation properties can be improved using heteronuclear multiple-quantum coherences (HMQCs) instead of heteronuclear single-quantum coherences (HSQCs) in the pulse sequence design. In slowly tumbling macromolecules, these HMQCs can exhibit significantly better transverse relaxation properties than HSQCs. However, HMQC-type experiments also exhibit resonance splittings due to multiple two- and three-bond homo- and heteronuclear scalar couplings. We describe here a family of pulsed- field gradient (PFG) HMQC-type triple-resonance experiments using simultaneous 1H and 13C constant-time (CT) periods to eliminate the t1 dependence of these scalar coupling effects. These simultaneous CT PFG-(HA)CANH and PFG-(HA)CA(CO)NH HMQC-type experiments exhibit sharper resonance line widths and often have better S/N ratios than the corresponding HSQC-type experiments. Results on proteins ranging in size from 6 to 30 kDa show average methine CH HMQC:HSQC enhancement factors of 1.10 ± 0.15, with about 40% of the cross peaks exhibiting better S/N ratios in the simultaneous CT-HMQC versions compared with the HSQC versions.

Keywords: Heteronuclear relaxation; Heteronuclear multiple-quantum coherence; Bovine pancreatic trypsin inhibitor;; Bovine pancreatic ribonuclease A; Influenza virus A NS-1 protein; BPTI–trypsin complex

Language: English

Document Type: Regular paper

Affiliations: 1: Center for Advanced Biotechnology and Medicine and Department of Molecular Biology and Biochemistry, Rutgers University, 679 Hoes Lane, Piscataway, NJ 08854-5638, U.S.A.

Publication date: 1997-01-01

• In this: publication
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• In this Subject: Anatomy & Physiology ,  Analytical Chemistry ,  Internal Medicine
• By this author: Swapna G.V.T. ;  Rios C.B. ;  Shang Z. ;  Montelione G.T.

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