title: NMR solution structure determination of large RNA-protein complexes.
abstract: Structure determination of RNA-protein complexes is essential for our understanding of the multiple layers of RNA-mediated posttranscriptional regulation of gene expression. Over the past 20years, NMR spectroscopy became a key tool for structural studies of RNA-protein interactions. Here, we review the progress being made in NMR structure determination of large ribonucleoprotein assemblies. We discuss approaches for the design of RNA-protein complexes for NMR structural studies, established and emerging isotope and segmental labeling schemes suitable for large RNPs and how to gain distance restraints from NOEs, PREs and EPR and orientational information from RDCs and SAXS/SANS in such systems. The new combination of NMR measurements with MD simulations and its potential will also be discussed. Application and combination of these various methods for structure determination of large RNPs will be illustrated with three large RNA-protein complexes (>40kDa) and other interesting complexes determined in the past six and a half years.
authors.affiliation: Central European Institute of Technology, Masaryk University, Kamenice 753/5, 62500 Brno, Czech Republic.
authors.forename: Deepak Kumar
authors.initials: DK
authors.lastname: Yadav
authors.affiliation: Central European Institute of Technology, Masaryk University, Kamenice 753/5, 62500 Brno, Czech Republic. Electronic address: peter.lukavsky@ceitec.muni.cz.
authors.forename: Peter J
authors.initials: PJ
authors.lastname: Lukavsky
mesh_terms: D001665:Binding Sites; D007553:Isotope Labeling / Q000379:methods; D007554:Isotopes / Q000737:chemistry; D008958:Models, Molecular; D019906:Nuclear Magnetic Resonance, Biomolecular / Q000379:methods*; D009690:Nucleic Acid Conformation; D011487:Protein Conformation; D011506:Proteins / Q000737:chemistry*; D012313:RNA / Q000737:chemistry*; D012261:Ribonucleoproteins / Q000737:chemistry*
chemical_list: D007554:Isotopes; D011506:Proteins; D012261:Ribonucleoproteins; D012313:RNA
keywords: Biomolecular NMR; Integrated structural biology; Isotope labeling; NMR assignments; NMR structure determination; RNA-protein complex; RNA-protein interaction