SOLUTION STRUCTURE OF RNASE P RNA (M1 RNA) P4 STEM OLIGORIBONUCLEOTIDE COMPLEXED WITH COBALT (III) HEXAMINE, NMR, MINIMIZED AVERAGE STRUCTURE
Solution structure and metal-ion binding of the P4 element from bacterial RNase P RNA.
1F6X shows the structure of the free wildtype P4 stem oligoribonucleotide
1F6Z shows the structure of the free C70U mutant P4 stem oligoribonucleotide
enzymatically synthesized from DNA ologonucleotide template by T7 RNA polymerase
This structure was determined using standard 2D homonuclear techniques and 13C and 31P heteronuclear techniques at natural abundance. Intermolecular NOE crosspeaks between RNA protons and cobalt (III) hexammine protons and derived intermolecular distance constraints were used to determine the site of cobalt (III) hexammine binding in the complex structure
The average structure is based on superposition of 11 converged structures after refinement. The average RMS deviation between the ensemble and the average structure is 1.7 Angstrom. A total of 266 NOE-derived intramolecular distance constraints, 164 dihedral restraint and 48 distance restraints from hydrogen bonds were used in the refinement. 12 NOE derived intermolecular distance constraints were used to localize the bound cobalt (III) hexammine
restrained molecular dynamics and simulated annealing
2 mM P4 RNA; 100 mM sodium chloride; 10 mM phosphate buffer; 90% H2O, 10% D2O
2 mM P4 RNA; 100 mM sodium chloride; 10 mM phosphate buffer; 99.96% D2O
2 mM P4 RNA; 100 mM sodium chloride; 10 mM magnesium chloride; 10 mM phosphate buffer; 90% H2O, 10% D2O
2 mM P4 RNA; 100 mM sodium chloride; 3 mM hexammine cobalt chloride; 10 mM phosphate buffer; 90% H2O, 10% D2O
RIBONUCLEASE P, RIBOZYME, TRANSFER RNA PROCESSING, P4 STEM, METAL BINDING SITE, METAL COMPLEX, COBALT (III) HEXAMMINE COMPLEX, RNA