Correlation of the guanosine exchangeable and nonexchangeable base protons in13C-/15N-labeled RNA with an HNC-TOCSY-CH experiment

✍ Scribed by Jean-Pierre Simorre; Grant R. Zimmermann; Luciano Mueller; Arthur Pardi

• Publisher: Springer Netherlands
• Year: 1996
• Tongue: English
• Weight: 416 KB
• Volume: 7
• Category: Article
• ISSN: 0925-2738
• DOI: 10.1007/bf00203825

✦ Synopsis

A triple resonance HNC-TOCSY-CH experiment is described for correlating the guanosine imino proton and H8 resonances in 13C-/15N-labeled RNAs. Sequential assignment of the exchangeable imino protons in Watson-Crick base pairs is generally made independently of the assignment of the nonexchangeable base protons. This H(NC)-TOCSY-(C)H experiment makes it possible to unambiguously link the assignment of the guanosine H8 resonances with sequential assignment of the guanosine imino proton resonances. 2D H(NC)-TOCSY-(C)H spectra are presented for two isotopically labeled RNAs, a 30-nucleotide lead-dependent ribozyme known as the leadzyme, and a 48-nucleotide hammerhead ribozyme-RNA substrate complex. The results obtained on these two RNAs demonstrate that this HNC-TOCSY-CH experiment is an important tool for resonance assignment of isotopically labeled RNAs.