title: Sparse labeling of proteins: structural characterization from long range constraints.
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.
authors.affiliation: Complex Carbohydrate Research Center, University of Georgia, Athens, GA 30602, United States. Electronic address: jpresteg@ccrc.uga.edu.
authors.forename: James H
authors.initials: JH
authors.lastname: Prestegard
authors.affiliation: The Howard Hughes Medical Institute, Dept. Biochem. & Biophys., Univ. Calif. San Francisco, San Francisco, CA 94158, United States.
authors.forename: David A
authors.initials: DA
authors.lastname: Agard
authors.affiliation: Complex Carbohydrate Research Center, University of Georgia, Athens, GA 30602, United States.
authors.forename: Kelley W
authors.initials: KW
authors.lastname: Moremen
authors.affiliation: The Howard Hughes Medical Institute, Dept. Biochem. & Biophys., Univ. Calif. San Francisco, San Francisco, CA 94158, United States.
authors.forename: Laura A
authors.initials: LA
authors.lastname: Lavery
authors.affiliation: Complex Carbohydrate Research Center, University of Georgia, Athens, GA 30602, United States.
authors.forename: Laura C
authors.initials: LC
authors.lastname: Morris
authors.affiliation: Complex Carbohydrate Research Center, University of Georgia, Athens, GA 30602, United States.
authors.forename: Kari
authors.initials: K
authors.lastname: Pederson
mesh_terms: D008958:Models, Molecular; D008968:Molecular Conformation; D019906:Nuclear Magnetic Resonance, Biomolecular / Q000379:methods*; D017434:Protein Structure, Tertiary; D011506:Proteins / Q000737:chemistry*; D013113:Spin Labels
chemical_list: D011506:Proteins; D013113:Spin Labels
keywords: Glycoprotein; Ligand docking; NMR assignments; Paramagnetic constraints; Protein NMR; Residual dipolar coupling; Sparse labeling