Long-range 15N-1H correlation experiments for histidine rings: Difference between revisions
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=== Varian === | === Varian === | ||
[[File:gNfhsqc His.tgz|gNfhsqc_His.tgz]]: long range 15N-1H HSQC package - tested with VnmrJ 2.1B | |||
This pulse sequence correlates aromatic H-C protons of His with ring nitrogens. Modified from BioPack <tt>gNfhsqc.c</tt> by adding 90-180-90 composite 180 pulses on 15N and changing decoupling options. | |||
Tarball contents: | |||
*vnmrsys/psglib/gNfhsqc_His.c | *vnmrsys/psglib/gNfhsqc_His.c | ||
*vnmrsys/templates/layout/gNfhsqc_His/ | *vnmrsys/templates/layout/gNfhsqc_His/ | ||
| Line 23: | Line 13: | ||
Installation: | Installation: | ||
*Save the <tt>gNfhsqc_His.tgz</tt> file into VnmrJ user home directory (e.g. <tt>/home/vnmr1</tt>). | |||
*Extract with <tt>tar xvzf gNfhsqc_His.tar</tt> - existing files will be overwritten! | |||
* | Default settings | ||
* | *JNH=22.75 Hz, to refocus 1J_NH couplings | ||
*15N carrier at 200 ppm | |||
*15N spectral width is 200 ppm | |||
*13C carrier at 125 ppm | |||
*15N decoupling during acquisition is off | |||
*13C 40 ppm WURST decoupling during acquisition | |||
*3-9-19 for water suppression | |||
Points to consider: | |||
*WURST decoupling produces strong side-bands, especially harmful if your protein has a His-tag. It is thus recommended to set 13C decoupling to GARP. | |||
*On some instruments 90-180-90 composite 15N pulses may be longer than the corresponding 3-9-19 delay, creating a negative delay in the pulse sequence (look for the '?' in 'dps'). In this case use traditional WATERGATE instead of 3-9-19 | |||
There is also the <tt>rna_WGgNhsqclr</tt> setup macro from BioPack. This macro loads the <tt>rna_WGgNhsqc.c</tt> pulsesequece with parameters for long-range correlations. | |||
Reference: [http://www.pubmedcentral.nih.gov/articlerender.fcgi?artid=2142369 J. G. Pelton, D. A. Torchia, N. D. Meadow and S. Roseman, ''Protein Sci'' 1993 2: 543-558] | Reference: [http://www.pubmedcentral.nih.gov/articlerender.fcgi?artid=2142369 J. G. Pelton, D. A. Torchia, N. D. Meadow and S. Roseman, ''Protein Sci'' 1993 2: 543-558] | ||
Latest revision as of 14:27, 7 October 2010
2D [15N, 1H] long-range HSQC for His rings
Varian
File:GNfhsqc His.tgz: long range 15N-1H HSQC package - tested with VnmrJ 2.1B
This pulse sequence correlates aromatic H-C protons of His with ring nitrogens. Modified from BioPack gNfhsqc.c by adding 90-180-90 composite 180 pulses on 15N and changing decoupling options.
Tarball contents:
- vnmrsys/psglib/gNfhsqc_His.c
- vnmrsys/templates/layout/gNfhsqc_His/
- vnmrsys/maclib/gNfhsqc_His
Installation:
- Save the gNfhsqc_His.tgz file into VnmrJ user home directory (e.g. /home/vnmr1).
- Extract with tar xvzf gNfhsqc_His.tar - existing files will be overwritten!
Default settings
- JNH=22.75 Hz, to refocus 1J_NH couplings
- 15N carrier at 200 ppm
- 15N spectral width is 200 ppm
- 13C carrier at 125 ppm
- 15N decoupling during acquisition is off
- 13C 40 ppm WURST decoupling during acquisition
- 3-9-19 for water suppression
Points to consider:
- WURST decoupling produces strong side-bands, especially harmful if your protein has a His-tag. It is thus recommended to set 13C decoupling to GARP.
- On some instruments 90-180-90 composite 15N pulses may be longer than the corresponding 3-9-19 delay, creating a negative delay in the pulse sequence (look for the '?' in 'dps'). In this case use traditional WATERGATE instead of 3-9-19
There is also the rna_WGgNhsqclr setup macro from BioPack. This macro loads the rna_WGgNhsqc.c pulsesequece with parameters for long-range correlations.
Reference: J. G. Pelton, D. A. Torchia, N. D. Meadow and S. Roseman, Protein Sci 1993 2: 543-558