Trp e1 and d1 assignment with NOESY

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Assignment of the Trp e1 and d1 is easily obtained with the [15N, 1H]-HSQC and simultaneous [1H, 1H]-NOESY


From b → d1 → e1

  • Prepare the simultaneous [1H, 1H]-NOESY in XEASY (ls, lc, lp).
  • Use sp (sp and type TRP in the box labeled fragment type and type 0 in the box labeled sequential range), es, se, gs to produce strips containing only peaks belonging to Trp residues.
  • Start in the strips containing the CB/HB peaks and look for peak around 7ppm. Move the HD1 peak marker vertically to this peak and then update the strip list (es, se, gs)
  • Change the orientation of the 3D spectrum to X(13C/15N), Y(1H NOESY), Z(1H detect) using pm and then gs to display the strips.
  • Move the HD1 peak marker horizontally (use eh, gr, gl, and za to expand, move, and zoom in as needed) to the correct position. Update the strips (es, se, gs), change back to the standard orientation [pm: X(13C/15N), Y(1H NOESY), Z(1H detect)], and display the strips again (gs).
  • From the newly adjusted HD1 strip, you should see the HE1 peak below (around 10ppm). Move the HE1 peak marker to this position and update the strips (es, se, gs).
  • Change the orientation of the 3D spectrum to X(13C/15N), Y(1H NOESY), Z(1H detect) using pm and then gs to display the strips.
  • Move the HE1 peak marker horizontally (use eh, gr, gl, and za to expand, move, and zoom in as needed) to the correct position. Update the strips (es, se, gs), change back to the standard orientation [pm: X(13C/15N), Y(1H NOESY), Z(1H detect)], and display the strips again (gs).
  • Confirm and adjust the positions of HD1 and HE1 if needed.
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