nightshift

A python library and command line program for plotting simulated 2D and 3D NMR spectra from assigned chemical shifts from the BMRB

Peak assignments are pulled using the BMRB API (https://github.com/uwbmrb/BMRB-API) and plotted with matplotlib. Spectra produced by this script do not always reflect reality, as many entries do not have 100% assignments. Be sure to know what assignments are contained in the entry beforehand.

Citation

If you find this code useful please cite our paper:

Fucci, I. J. and Byrd, R. A. nightshift: A Python program for plotting simulated NMR spectra from assigned chemical shifts from the Biological Magnetic Resonance Data Bank. Protein Sci. 2022, 31 (1), 63–74. https://doi.org/10.1002/pro.4181.

Installation

I recommend installing in a virtualenv to avoid any conflicts with your python installation. nightshift is available on the PyPI and can be installed with pip:

pip install nightshift

To install a local version downloaded from GitHub, use:

pip install /path/to/nightshift

Usage

You will need to know your protein of interest's BMRB entry number. You can use nightshift's search command to find entry numbers:

nightshift search ubiquitin

We'll use 4493 Solution structure of the designed hydrophobic core mutant of ubiquitin, 1D7 for our examples.

Basic spectra

Two major use cases are getting an idea what a amide or methyl spectrum of your protein would look like

To simulate 1H-15N HSQC spetra can be plotted using the --amide flag:

nightshift get 4493 --amide

To show Asn and Gln side chain amides on the spectrum pass the optional --sidechains flag:

nightshift get 4493 --amide --sidechains

A simulated 1H-13C HMQC spetra can be plotted using the --methyl flag, additionally providing the optional proS or proR will filter LV atoms by prochirality:

nightshift get 4493 --methyl --proS

All plots can be filtered based on residue type by passing the --residues or -r flag and providing one-letter amino acid codes. For instance an ILV methyl labeled spectrum can be plotted using:

nightshift get 4493 --methyl -r ILV

Custom correlations

For arbitrary correlations use the --custom flag followed by two atom names. Consider yourself warned that labeling schemes and/or experiments to produce these correlations may not (currently) exist. Atoms for custom correlations are specified using standard PDB atom names: H for amide proton, N for amide nitrogen, C for carbonyl carbon, CA for alpha carbon, HA for alpha proton and so on. For particular residues two or more atoms may exist at a position (i.e. CG for Val could be CG1 or CG2). To specify both CG1 and CG2 for Val pass CG:

nightshift get 4493 --custom CG CA -r V

or specify the full atom name to only get those atoms:

nightshift get 4493 --custom CG1 CA -r V

Two special atom names also exist for custom correlations: Hmethyl and Cmethyl. Which correspond to these atoms in MILVAT residues and are the same atoms selected by using the --methyl flag. The flags --proS and --proR can be used with Hmethyl and Cmethyl

| Residue | Hmethyl | Cmethyl | |---------|------------|---------| | Met | HE1 | CE | | Ile | HD11 | CD1 | | Leu | HD11, HD21 | CD1,CD2 | | Val | HG11, HG21 | CG1,CG2 | | Ala | HB1 | CB | | Thr | CG2 | HG21 |

This allows for correlations of methyl groups to any other atom to be plotted. For instance Cmethyl to CA:

nightshift get 4493 --custom Cmethyl CA -r ILV

Adding '-' or '+' and any number to the end of a custom atom name allows correlation to the i+/- num residue. For instance correlation of the CO of the i-1 residue to the amide N of the i residue:

nightshift get 4493 --custom C-1 N

3D correlations

The --custom option also allows for 3D correlations to be plotted. The matplotlib window will show 2D slices of the simulated spectrum. Scrolling the mouse wheel will switch between each slice:

nightshift get 4493 --custom H N CA

By default 16 slices are generated, this can be altered with the --slices option (i.e. slices 32 or slices 1 for a 2D projection). The --project parameter can take a value of 1, 2 or 3 which chooses which dimension to project on. For a 3D HNCA (NOTE: you must escape with double quotes):

nightshift get 4493 --custom "H N (CA CA-1)" --project 2 --slices 32

Plus and minus can also be used on 3D correlations:

nightshift get 4493 --custom HA N+1 CA+2 --project 1

Other options

By default plots are generated in matplotlib and are interactive. To save directly to an image file use the --output or -o flag and provide a file name and extension (.eps, .pdf, .pgf, .png, .ps, .raw, .rgba, .svg, and .svgz are all acceptable).

Formatting options include: - --showlegend to add a legend - --nolabels to remove the residue/atom name and numbers from the plot, also shows the legend - --offset to add a constant to the indices used by BMRB (to reflect the numbering you are used to) - --color takes a color such as red which will color all residues one color or a matplotlib colormap name such as viridis or tab20 which will color each residue with a differen color on the colormap

A csv file containing the label and chemical shifts of both atoms can be saved using the --csv flag and providing a file name:

nightshift get 4493 --methyl -r ILV --csv output.csv

this can be opened in other software to generate plots with different formatting. Also the auxiliary script plot_outputs.py can be used to overlay multiple spectra (i.e. different domains of the same protein or a protein complex).

This example is nonsense, but illustrates how it could be done. First, generate two output files:

nightshift get 4493 --amide --csv output1.csv

nightshift get 3433 --amide --csv output2.csv

Then plot their overlay:

nightshift open output1.csv output2.csv

If two labels are the same between the proteins (labels are residue type and sequence number), you can visualize tahe CSP using --showcsp, which will draw a line between the two. This works for 4493 and 3433, but is not very informative.

nightshift open output1.csv output2.csv --showcsp

Also accepts the --showlegend and --nolabels flags. --colors takes a list of color names or list of colormaps and works like --color for individual plots.

Interesting examples

• ILV methyl spectrum:

nightshift get 4493 --methyl -r ILV

• ILV methyl spectrum with proR LV:

nightshift get 4493 --methyl proR -r ILV

• Amide spectrum of only lysines and arginines:

nightshift get 4493 --amide -r KR

• 2D HMBC-HMQC (intra-residue methyl-methyl correlations):

nightshift get 4493 --custom Cmethyl Cmethyl -r LV

• 2D NCO:

nightshift get 4493 --custom C-1 N

• 2D CAN

nightshift get 4493 --custom "(CA CA-1) N" --label 1

• 3D HNCACB nightshift get 4493 --custom "H N (CA CA-1 CB CB-1)" --project 2 --label 3 --slices 32

• 3D 15N TOCSY HSQC nightshift get 4493 --custom "H H# N" --slices 64

• Arg/Lys side chain carbon correlations (a la Pritchard and Hansen, 2019)

nightshift get 4493 --custom CG CD -r R --csv 4493_arg.csv

nightshift get 4493 --custom CD CE -r K --csv 4493_lys.csv

nightshift open 4493_arg.csv 4493_lys.csv --showlegend

Transfering assignments to nmrDraw

I have only tested that this works for amide and methyl because of limitations with NMRpipe. It is possible other correllations will work fine. See the cs2pk.tcl documentation. There is a hacky work around where methyl spectra are considered HA/CA spectra. To generate the TALOS file required as input use the --talos flag: nightshift get 4493 --amide --talos 4493.tab

or

nightshift get 4493 --methyl -r ILV --talos 4493.tab

To transfer run the following for amide:

cs2pk.tcl -in 4493.tab -out sim.tab -type hn

and for methyl:

cs2pk.tcl -in <talos.tab> -out <sim.tab> -type haca

Compare against your spectrum visually (assuming the name of the file is test.ft2 and the peak list is test.tab):

specView.tcl -in test.ft2 -multiColor -peak -tab test.tab -pkLab ASS -ass sim.tab -outTab out.tab -xAssVar X_PPM -yAssVar Y_PPM

or automatically without an interface:

ipap.tcl -specName1 test.ft2 -inName1 test.tab -outName1 out.tab -assName sim.tab -xVar X_PPM -xAss X_PPM -yVar Y_PPM -yAss Y_PPM -single -ndim 2 -exit

See the documentation for specView.tcl and ipap.tcl for a full list of parameters.