Transverse relaxation optimized HCN experiment for nucleic acids: combining the advantages of TROSY and MQ spin evolution.
A three-dimensional MQ-TROSY-HCN pulse sequence is presented which provides intra-base and sugar-to-base correlations for 13C, 15N labeled nucleic acids (RNA, DNA). The experiment simultaneously exploits the favorable relaxation properties of 1H-13C multiple quantum coherence for sugar carbons and of 13C TROSY-type spin evolution for base carbons. MQ-TROSY-HCN thus combines the advantages of MQ-HCN for sugar-to-base and TROSY-HCN for intra-base correlations in a single experiment. In addition, two slightly different implementations of the MQ-TROSY-HCN experiment ensure optimal performance for small and larger oligonucleotides, respectively. The advantages of the MQ-TROSY-HCN experiment compared to the best previous implementations of HCN are demonstrated for a 33 nucleotide RNA aptamer.
Brutscher
B
B
Institut de Biologie Structurale, Jean-Pierre Ebel CNRS-CEA, Grenoble, France. Bernhard.Brutscher@ibs.fr
Simorre
J P
JP
D001483:Base Sequence; D002241:Carbohydrates / Q000737:chemistry; D006856:Hydrogen Cyanide / Q000737:chemistry*; D019906:Nuclear Magnetic Resonance, Biomolecular / Q000379:methods*; D009690:Nucleic Acid Conformation; D009696:Nucleic Acids / Q000737:chemistry*; D009843:Oligoribonucleotides / Q000737:chemistry; D011522:Protons; D012313:RNA / Q000737:chemistry
D002241:Carbohydrates; D009696:Nucleic Acids; D009843:Oligoribonucleotides; D011522:Protons; D006856:Hydrogen Cyanide; D012313:RNA