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J Magn Reson. 2014 Apr;241:32-40. doi: 10.1016/j.jmr.2013.12.012.

Sparse labeling of proteins: structural characterization from long range constraints.

Author information

1
Complex Carbohydrate Research Center, University of Georgia, Athens, GA 30602, United States. Electronic address: jpresteg@ccrc.uga.edu.
2
The Howard Hughes Medical Institute, Dept. Biochem. & Biophys., Univ. Calif. San Francisco, San Francisco, CA 94158, United States.
3
Complex Carbohydrate Research Center, University of Georgia, Athens, GA 30602, United States.

Abstract

Structural characterization of biologically important proteins faces many challenges associated with degradation of resolution as molecular size increases and loss of resolution improving tools such as perdeuteration when non-bacterial hosts must be used for expression. In these cases, sparse isotopic labeling (single or small subsets of amino acids) combined with long range paramagnetic constraints and improved computational modeling offer an alternative. This perspective provides a brief overview of this approach and two discussions of potential applications; one involving a very large system (an Hsp90 homolog) in which perdeuteration is possible and methyl-TROSY sequences can potentially be used to improve resolution, and one involving ligand placement in a glycosylated protein where resolution is achieved by single amino acid labeling (the sialyltransferase, ST6Gal1). This is not intended as a comprehensive review, but as a discussion of future prospects that promise impact on important questions in the structural biology area.

KEYWORDS:

Glycoprotein; Ligand docking; NMR assignments; Paramagnetic constraints; Protein NMR; Residual dipolar coupling; Sparse labeling

PMID:
24656078
PMCID:
PMC3964372
DOI:
10.1016/j.jmr.2013.12.012
[Indexed for MEDLINE]
Free PMC Article

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