title: Automated evaluation of chemical shift perturbation spectra: New approaches to quantitative analysis of receptor-ligand interaction NMR spectra.
abstract: This paper presents new methods designed for quantitative analysis of chemical shift perturbation NMR spectra. The methods automatically trace the displacements of cross peaks between a perturbed test spectrum and the reference spectrum (or among a series of titration spectra), and measure the changes of chemical shifts, heights, and widths of the altered peaks. The methods are primary aimed at the (1)H-(15)N HSQC spectra of relatively small proteins (<15 kDa) assuming fast exchange between free and ligand-bound states on the chemical shift time scale, or for comparing spectra of free and fully bound states in the slow exchange situation. Using the (1)H-(15)N HSQC spectra from a titration experiment of the 74-residue Pex13p SH3 domain with a Pex14p peptide ligand (14 residues, K (d)= approximately 40 microM), we demonstrate the scope and limits of our automatic peak tracing (APET) algorithm for efficient scoring of high-throughput SAR by NMR type HSQC spectra, and progressive peak tracing (PROPET) algorithm for detailed analysis of ligand titration spectra. Simulated spectra with low signal-to-noise ratios (S/N ranged from 20 to 1) were used to demonstrate the reliability and reproducibility of the results when dealing with poor quality spectra. These algorithms have been implemented in a new software module, FELIX-Autoscreen, for streamlined processing, analysis and visualization of SAR by NMR and other high-throughput receptor/ligand interaction experiments.
authors.affiliation: Accelrys Inc., 9685 Scranton Road, San Diego, CA 92121, U.S.A.
authors.forename: Chen
authors.initials: C
authors.lastname: Peng
authors.forename: Stephen W
authors.initials: SW
authors.lastname: Unger
authors.forename: Fabian V
authors.initials: FV
authors.lastname: Filipp
authors.forename: Michael
authors.initials: M
authors.lastname: Sattler
authors.forename: Sándor
authors.initials: S
authors.lastname: Szalma
mesh_terms: D000465:Algorithms; D003627:Data Interpretation, Statistical; D008024:Ligands; D009682:Magnetic Resonance Spectroscopy / Q000379:methods*; D011956:Receptors, Cell Surface / Q000737:chemistry* / Q000378:metabolism; D018160:Receptors, Cytoplasmic and Nuclear / Q000737:chemistry* / Q000378:metabolism
chemical_list: D008024:Ligands; D011956:Receptors, Cell Surface; D018160:Receptors, Cytoplasmic and Nuclear