Amino acid type-selective experiments can help to remove ambiguities in automated assignment procedures for 15 N/ 13 C-labeled proteins. Here we present five triple-resonance experiments that yield amino acid type-selective 1 H-15 N correlations for aromatic amino acids. Four of the novel experiment
MUSIC, Selective Pulses, and Tuned Delays: Amino Acid Type-Selective 1H–15N Correlations, II
✍ Scribed by Mario Schubert; Hartmut Oschkinat; Peter Schmieder
- Publisher
- Elsevier Science
- Year
- 2001
- Tongue
- English
- Weight
- 177 KB
- Volume
- 148
- Category
- Article
- ISSN
- 1090-7807
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✦ Synopsis
Amino acid type-selective experiments help to remove ambiguities in either manual or automated assignment procedures. Here we present modified triple-resonance experiments that yield amino acid type-selective (1)H-(15)N correlations. They are based on the MUSIC coherence transfer scheme which replaces the initial INEPT transfer and is selective for XH(2) or XH(3) (where X is either (15)N or (13)C). Signals of the desired amino acid types are thus selected based on the topology of the side chain. MUSIC is combined with selective pulses and carefully tuned delays to create experiments for Ser (S-HSQC); Val, Ile, and Ala (VIA-HSQC); Leu and Ala (LA-HSQC); Asp, Asn, and Gly (DNG-HSQC), as well as Glu, Gln, and Gly (EQG-HSQC). The new experiments are recorded as two-dimensional spectra and their performance is demonstrated by their application to two protein domains of 83 and 115 residues.
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Amino acid type-selective triple-resonance experiments can be of great help for the assignment of protein spectra, since they help to remove ambiguities in either manual or automated assignment procedures. Here, modified triple-resonance experiments that yield amino acid type-selective 1 H-15 N corr