¹⁵N/¹³C_m-HSQC and ¹⁵N/¹³C_m-TROSY/HSQC

Pulse sequences for ¹⁵N/¹³C_m-HSQC and
¹⁵N/¹³C_m-TROSY/HSQC with ¹⁵N gradient selection.

Examples on  uniformly labeled ¹⁵N and fractional (~10%) ¹³C SH3 domain free in solution (red) and in 1:2 SH3:peptide (black)

Reference:

P. Würtz, O. Aitio, M. Hellman and P. Permi:
Simultaneous Detection of Amide and Methyl Correlations Using Time Shared NMR Experiment: Application to Binding Epitope Mapping
J. Biomol. NMR (2007); 39(2):97-105.

Download and installation

Simultaneous Detection of Amide and Methyl Correlations Using Time Shared NMR Experiment.
Download complete package: ts15n13chsqc.tgz

Installation instructions:

15N/¹³C_m-HSQC and ¹⁵N/¹³C_m-TROSY/HSQC pulse sequences for Varian Unity Inova. Tested with Vnmr 6.1c.

Please read the disclaimer before using the sequence. For usage/processing refer to the manual page.

Installation setup

1. Login at the user account in which the installation should be carried out.
2. Save the ts15n13c_hsqc.tgz file at a location with write permission for the user who is carrying out the installation.
3. Unpack the file with the command: tar xzvf ts15n13c_hsqc.tgz
4. Copy the files to the corresponding vnmrsys directories

Running and processing experiments

1. Start with an ¹⁵N HSQC experiment.
2. Load the parameter file.
3. Run the gNCm_hsqc macro or gNCm_trosyhsqcfor setting up the experiment.
4. Set relevant parameters such as pulse widths and power levels (specified in manual files).
5. Adjust receiver gain and start experiment.
6. Standard processing by wft2da (¹⁵N) and wft_NCm (¹³C) macroes.
7. ¹³C referecing in standard manner. ¹⁵N axis referecing by NC_scalenh followed by standard referencing.

For questions or requests please contact Dr. Perttu Permi, perttu.permihelsinki.fi