title: Reduced dimensionality (4,3)D-HN(C)NH for rapid assignment of 1H(N)-15N HSQC peaks in proteins: an analytical tool for protein folding, proteomics, and drug discovery programs.
abstract: While nuclear magnetic resonance (NMR) has had commendable success in atomic-level investigation of folded proteins, intrinsically unfolded and partially folded proteins have always posed a great challenge, because of poor chemical shift dispersions. We present here a reduced-dimensionality-based NMR triple resonance pulse sequence, (4,3)D-HN(C)NH, which not only helps to disperse the peaks further by combining (15)N and amide (1)H chemical shifts, but also directly establishes correlations between (1)H(i)(N), (15)N(i), (1)H(i+1)(N), and (15)N(i+1) spins along the F(1)-F(3) planes. The F(2)-F(3) projection planes of this experiment provide unique identification of the check points in amide resonances. An assignment strategy derived by combining information along the F(1)-F(3) planes and in the F(2)-F(3) projection planes of the experiment has been presented and shown to be very useful for both intrinsically disordered/unfolded proteins and folded protein alike. The experiment and the protocol would be valuable for protein folding, proteomics, and drug discovery programs.
authors.affiliation: Department of Chemical Sciences, Tata Institute of Fundamental Research, Colaba, Mumbai, India.
authors.forename: Jithender G
authors.initials: JG
authors.lastname: Reddy
authors.forename: Ramakrishna V
authors.initials: RV
authors.lastname: Hosur
mesh_terms: D002247:Carbon Isotopes / Q000737:chemistry; D003459:Crystallins / Q000737:chemistry* / Q000378:metabolism; D055808:Drug Discovery; D006801:Humans; D006859:Hydrogen / Q000737:chemistry; D009587:Nitrogen Isotopes / Q000737:chemistry; D019906:Nuclear Magnetic Resonance, Biomolecular; D017510:Protein Folding; D040901:Proteomics; D025842:SUMO-1 Protein / Q000737:chemistry* / Q000378:metabolism
chemical_list: D002247:Carbon Isotopes; D003459:Crystallins; D009587:Nitrogen Isotopes; D025842:SUMO-1 Protein; D006859:Hydrogen