title: Solution NMR Experiment for Measurement of (15)N-(1)H Residual Dipolar Couplings in Large Proteins and Supramolecular Complexes.
abstract: NMR residual dipolar couplings (RDCs) are exquisite probes of protein structure and dynamics. A new solution NMR experiment named 2D SE2 J-TROSY is presented to measure N-H RDCs for proteins and supramolecular complexes in excess of 200 kDa. This enables validation and refinement of their X-ray crystal and solution NMR structures and the characterization of structural and dynamic changes occurring upon complex formation. Accurate N-H RDCs were measured at 750 MHz (1)H resonance frequency for 11-mer 93 kDa (2)H,(15)N-labeled Trp RNA-binding attenuator protein tumbling with a correlation time τc of 120 ns. This is about twice as long as that for the most slowly tumbling system, for which N-H RDCs could be measured, so far, and corresponds to molecular weights of ∼200 kDa at 25 °C. Furthermore, due to the robustness of SE2 J-TROSY with respect to residual (1)H density from exchangeable protons, increased sensitivity at (1)H resonance frequencies around 1 GHz promises to enable N-H RDC measurement for even larger systems.
authors: Department of Chemistry and ‡Department of Biological Sciences, State University of New York at Buffalo , Buffalo, New York 14260, United States.
authors: Alexander
authors: A
authors: Eletsky
authors: Department of Chemistry and ‡Department of Biological Sciences, State University of New York at Buffalo , Buffalo, New York 14260, United States.
authors: Surya V S R K
authors: SV
authors: Pulavarti
authors: Department of Chemistry and ‡Department of Biological Sciences, State University of New York at Buffalo , Buffalo, New York 14260, United States.
authors: Victor
authors: V
authors: Beaumont
authors: Department of Chemistry and ‡Department of Biological Sciences, State University of New York at Buffalo , Buffalo, New York 14260, United States.
authors: Paul
authors: P
authors: Gollnick
authors: Department of Chemistry and ‡Department of Biological Sciences, State University of New York at Buffalo , Buffalo, New York 14260, United States.
authors: Thomas
authors: T
authors: Szyperski
mesh_terms: D001426:Bacterial Proteins / Q000737:chemistry; D009682:Magnetic Resonance Spectroscopy / Q000379:methods*; D008958:Models, Molecular; D008970:Molecular Weight; D011487:Protein Conformation; D011506:Proteins / Q000737:chemistry*; D016601:RNA-Binding Proteins / Q000737:chemistry; D012996:Solutions; D014157:Transcription Factors / Q000737:chemistry
chemical_list: D001426:Bacterial Proteins; C079092:MtrB protein, Bacteria; D011506:Proteins; D016601:RNA-Binding Proteins; D012996:Solutions; D014157:Transcription Factors