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Communication

Rapid Data Collection for Protein Structure Determination by NMR Spectroscopy

Supporting Info

Yingqi Xu, Dong Long, and Daiwen Yang*

Department of Biological Sciences, National University of Singapore, 14 Science Drive 4, Singapore 117543

J. Am. Chem. Soc., 2007, 129 (25), pp 7722–7723

DOI: 10.1021/ja071442e

Publication Date (Web): May 31, 2007

In papers with more than one author, the asterisk indicates the name of the author to whom inquiries about the paper should be addressed.

, dbsydw@nus.edu.sg

Abstract

A new approach is proposed to rapidly determine solution structures of small and medium-sized ¹³C,¹⁵N-labeled proteins, using a minimal number of experiments, such as 4D time-shared ¹³C/¹⁵N, ¹³C/¹⁵N-edited NOESY, 3D HNCA, and 3D MQ-(H)CCH-TOCSY. When the data are recorded on a 500 MHz spectrometer without using sparse sampling techniques, the experimental times were minimized to 2.5, 8.5, and 96 h for the HNCA, MQ-(H)CCH-TOCSY, and NOESY, respectively. The time-shared 4D NOESY contains four sub-spectra, one HC-NOESY-CH, one HC-NOESY-NH, one HN-NOESY-CH, and one HN-NOESY-NH. Sequence-specific assignment of backbone and side-chain resonances is achieved from the 3D spectra and the HC-NOESY-NH spectrum. Good initial structures can be quickly calculated using many unambiguous distance restraints that are easily obtained from the NOESY sub-spectra. The structures can be refined with more restraints assigned based on the initial structures. The approach has been demonstrated on ubiquitin (76 residues), liver fatty acid binding protein (129 residues, its NMR assignment and solution structure are presented here for the first time), and a cell−cell adhesion protein (214 residues). The approach will facilitate high throughput structure determination by NMR.

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