title: [13C]-constant-time [15N,1H]-TROSY-HNCA for sequential assignments of large proteins.
abstract: The greatly improved sensitivity resulting from the use of TROSY during 15N evolution and amide proton acquisition enables the recording of HNCA spectra of large proteins with constant-time 13C alpha evolution. In [13C]-ct-[15N,1H]-TROSY-HNCA experiments with a 2H/13C/15N-labeled 110 kDa protein, 7,8-dihydroneopterin aldolase from Staphylococcus aureus, nearly all correlation peaks seen in the [15N,1H]-TROSY-HNCA spectrum were also detected. The improved resolution in the 13C dimension then enabled a significant number of sequential assignments that could not be obtained with [15N,1H]-TROSY-HNCA without [13C]-constant-time period.
authors.affiliation: Institut für Molekularbiologie und Biophysik, Eidgenössische Technische Hochschule Hönggerberg, Zürich, Switzerland.
authors.forename: M
authors.initials: M
authors.lastname: Salzmann
authors.forename: K
authors.initials: K
authors.lastname: Pervushin
authors.forename: G
authors.initials: G
authors.lastname: Wider
authors.forename: H
authors.initials: H
authors.lastname: Senn
authors.forename: K
authors.initials: K
authors.lastname: Wüthrich
mesh_terms: D000446:Aldehyde-Lyases / Q000737:chemistry*; D000577:Amides; D002247:Carbon Isotopes; D003903:Deuterium; D006859:Hydrogen; D009587:Nitrogen Isotopes; D019906:Nuclear Magnetic Resonance, Biomolecular / Q000379:methods; D011487:Protein Conformation; D011506:Proteins / Q000737:chemistry*; D012680:Sensitivity and Specificity; D013211:Staphylococcus aureus / Q000201:enzymology; D013997:Time Factors
chemical_list: D000577:Amides; D002247:Carbon Isotopes; D009587:Nitrogen Isotopes; D011506:Proteins; D006859:Hydrogen; D003903:Deuterium; D000446:Aldehyde-Lyases; C020044:dihydroneopterin aldolase