title: Backbone resonance assignment in large protonated proteins using a combination of new 3D TROSY-HN(CA)HA, 4D TROSY-HACANH and 13C-detected HACACO experiments.
abstract: A new method for backbone resonance assignment suitable for large proteins with the natural (1)H isotope content is proposed based on a combination of the most sensitive TROSY-type triple-resonance experiments. These techniques include TROSY-HNCO, (13)C'-detected 3D multiple-quantum HACACO and the newly developed 3D TROSY multiple-quantum-HN(CA)HA and 4D TROSY multiple-quantum-HACANH experiments. The favorable relaxation properties of the multiple-quantum coherences, signal detection using the (13)C' antiphase coherences, and the use of TROSY optimize the performance of the proposed set of experiments for application to large protonated proteins. The method is demonstrated with the 44 kDa uniformly (15)N,(13)C-labeled and fractionally (35%) deuterated trimeric B. Subtilis Chorismate Mutase and is suitable for proteins with large correlation times but a relatively small number of residues, such as membrane proteins embedded in micelles or oligomeric proteins.
authors.affiliation: Laboratorium für Physikalische Chemie, Eidgenössische Technische Hochschule Hönggerberg, CH-8093 Zürich, Switzerland.
authors.forename: Kaifeng
authors.initials: K
authors.lastname: Hu
authors.forename: Alexander
authors.initials: A
authors.lastname: Eletsky
authors.forename: Konstantin
authors.initials: K
authors.lastname: Pervushin
mesh_terms: D002247:Carbon Isotopes; D009682:Magnetic Resonance Spectroscopy / Q000379:methods*; D008962:Models, Theoretical; D009587:Nitrogen Isotopes; D011487:Protein Conformation; D011506:Proteins / Q000737:chemistry*; D011522:Protons; D011789:Quantum Theory; D012680:Sensitivity and Specificity
chemical_list: D002247:Carbon Isotopes; D009587:Nitrogen Isotopes; D011506:Proteins; D011522:Protons