Measuring protein self-diffusion in protein-protein mixtures using a pulsed gradient spin-echo technique with WATERGATE and isotope filtering.
Here we report a modified pulsed gradient spin-echo (PGSTE) pulse sequence to measure diffusion coefficients. This approach incorporates WATERGATE combined with isotopic filtering into a standard PGSTE experiment. Doing this eliminates much of the disadvantages from the combination of diffusion encoding and heteronuclear selection intervals and allows for facile modification of the diffusion pulse sequence with flexibility of the time period between RF pulses. The new diffusion pulse sequence is demonstrated using an 15N-labeled peptide and an 15N-labeled protein in a mixture with a protein of similar size.
Nesmelova
Irina V
IV
Department of Biochemistry, Molecular Biology and Biophysics, University of Minnesota, Minneapolis, MN 55455, USA.
Idiyatullin
Djaudat
D
Mayo
Kevin H
KH
D000465:Algorithms; D045424:Complex Mixtures / Q000032:analysis* / Q000737:chemistry*; D004058:Diffusion; D009587:Nitrogen Isotopes; D019906:Nuclear Magnetic Resonance, Biomolecular / Q000379:methods*; D010455:Peptides / Q000032:analysis / Q000737:chemistry; D011506:Proteins / Q000032:analysis* / Q000737:chemistry*; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D012815:Signal Processing, Computer-Assisted; D012996:Solutions
D045424:Complex Mixtures; D009587:Nitrogen Isotopes; D010455:Peptides; D011506:Proteins; D012996:Solutions