title: NMR resonance assignments for sparsely 15N labeled proteins.
abstract: For larger proteins, and proteins not amenable to expression in bacterial hosts, it is difficult to deduce structures using NMR methods based on uniform (13)C, (15)N isotopic labeling and observation of just nuclear Overhauser effects (NOEs). In these cases, sparse labeling with selected (15)N enriched amino acids and extraction of a wider variety of backbone-centered structural constraints is providing an alternate approach. A limitation, however, is the absence of resonance assignment strategies that work without uniform (15)N, (13)C labeling or preparation of numerous samples labeled with pairs of isotopically labeled amino acids. In this paper an approach applicable to a single sample prepared with sparse (15)N labeling in selected amino acids is presented. It relies on correlation of amide proton exchange rates, measured from data on the intact protein and on digested and sequenced peptides. Application is illustrated using the carbohydrate binding protein, Galectin-3. Limitations and future applications are discussed.
authors.affiliation: Complex Carbohydrate Research Center, University of Geogia, Athens, GA 30602-4712, USA.
authors.forename: Lianmei
authors.initials: L
authors.lastname: Feng
authors.forename: Han-Seung
authors.initials: HS
authors.lastname: Lee
authors.forename: James H
authors.initials: JH
authors.lastname: Prestegard
mesh_terms: D000577:Amides / Q000737:chemistry; D000595:Amino Acid Sequence; D003903:Deuterium; D037502:Galectin 3 / Q000737:chemistry* / Q000378:metabolism; D008969:Molecular Sequence Data; D009584:Nitrogen / Q000032:analysis* / Q000737:chemistry; D009587:Nitrogen Isotopes; D019906:Nuclear Magnetic Resonance, Biomolecular / Q000379:methods*; D010455:Peptides / Q000737:chemistry; D010649:Phenylalanine / Q000737:chemistry; D017510:Protein Folding; D017434:Protein Structure, Tertiary; D011522:Protons
chemical_list: D000577:Amides; D037502:Galectin 3; D009587:Nitrogen Isotopes; D010455:Peptides; D011522:Protons; D010649:Phenylalanine; D003903:Deuterium; D009584:Nitrogen