Details

Autor(en) / Beteiligte

• Long, Dong
• Delaglio, Frank
• Sekhar, Ashok
• Kay, Lewis E.

Titel

Probing Invisible, Excited Protein States by Non-Uniformly Sampled Pseudo-4D CEST Spectroscopy

Ist Teil von

• Angewandte Chemie, 2015-09, Vol.127 (36), p.10653-10657

Ort / Verlag

Weinheim: WILEY-VCH Verlag

Erscheinungsjahr

2015

Quelle

Wiley-Blackwell Full Collection

Beschreibungen/Notizen

• Chemical exchange saturation transfer (CEST) NMR spectroscopy is a powerful tool for studies of slow timescale protein dynamics. Typical experiments are based on recording a large number of 2D data sets and quantifying peak intensities in each of the resulting planes. A weakness of the method is that peaks must be resolved in 2D spectra, limiting applications to relatively small proteins. Resolution is significantly improved in 3D spectra but recording uniformly sampled data is time‐prohibitive. Here we describe non‐uniformly sampled HNCO‐based pseudo‐4D CEST that provides excellent resolution in reasonable measurement times. Data analysis is done through fitting in the time domain, without the need of reconstructing the frequency dimensions, exploiting previously measured accurate peak positions in reference spectra. The methodology is demonstrated on several protein systems, including a nascent form of superoxide dismutase that is implicated in neurodegenerative disease. Leistungsfähiges NMR‐Werkzeug: Ein HNCO‐basiertes Pseudo‐4D‐CEST‐Experiment mit nicht‐uniformem Sampling wird beschrieben, das Datensätze mit exzellenter Auflösung in praktikablen Aufnahmezeiten erzeugt. Die Datenanalyse wird in der Zeitdomäne durchgeführt und nutzt Vorabkenntnisse der Signallagen aus Referenzspektren.