A general strategy for the assignment of aliphatic side-chain resonances of uniformly 13C,15N-labeled large proteins

Yingqi Xu; Zhi Lin; Chien Ho; Daiwen Yang

doi:10.1021/ja053539b

A general strategy for the assignment of aliphatic side-chain resonances of uniformly 13C,15N-labeled large proteins

J Am Chem Soc. 2005 Aug 31;127(34):11920-1. doi: 10.1021/ja053539b.

Authors

Yingqi Xu¹, Zhi Lin, Chien Ho, Daiwen Yang

Affiliation

¹ Department of Biological Sciences, National University of Singapore, 14 Science Drive 4, Singapore 117543.

PMID: 16117513
DOI: 10.1021/ja053539b

Abstract

A general strategy is proposed to assign aliphatic side-chain resonances of large 13C,15N-labeled proteins without deuteration, using 4D 13C,15N-edited NOESY and MQ-(H)CCH-TOCSY experiments on the basis of prior assignments of backbone and 13Cbeta resonances. The strategy has been tested on a 214 residue protein (DdCAD-1) and applied to a chain-selectively 13C,15N-labeled hemoglobin (65 kDa). About 96 and 80% aliphatic side-chain spins in DdCAD-1 and hemoglobin have been assigned, respectively. The strategy proposed here will be very useful for the structure determination and dynamics characterization of large proteins by NMR.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Carbon Isotopes / chemistry*
Hydrocarbons, Acyclic / chemistry*
Molecular Weight
Nitrogen Isotopes / chemistry*
Nuclear Magnetic Resonance, Biomolecular / methods
Protein Conformation
Proteins / chemistry*

Substances

Carbon Isotopes
Hydrocarbons, Acyclic
Nitrogen Isotopes
Proteins