Normal mode analysis (Phenylketonuria)

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Summary

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Normal Mode Analysis

Lab journal
For the analysis, we used the pdb structure 1J8U from the task before (Task 9 - Structure-based mutation analysis).

WEBnm@

Mode 7

Mode 7 captured of WEBnm@. The PAH structure (pdb 1j8u) is shown in transperent green, the ligands in red (H4B: C₉H₁₅N₅O₃) and orange (Fe₂). The blue doted lines represent the movement of the structure.

</figure> <figure id="web_plot7">

'Normalized squared displacement per residue for mode 7 to 12. The x-axis represent the residue number in the sequence, whereby 0 is the first of the structure sequence, which is residue 103 in the FASTA-sequence of P00439. the y-axis shows the normalized squared displacement.

</figure>

This mode seems to have a high movement as the blue doted lines in <xr id="web_mode7"/> are relatively long. Furthermore they seem to open and close on all sites the same, which gives the imagine of a pump and not a twist. We additionally looked at the movement provided by the webserver, where the "breathing" rhythm could be seen, too. This also can be viewed in <xr id="web_plot7"/>. Between residues 20 and 30 there is a high atomic displacement which also can be seen for residues 240 to 300.

Mode 8

Mode 8 captured of WEBnm@. The PAH structure (pdb 1j8u) is shown in transperent green, the ligands in red (H4B: C₉H₁₅N₅O₃) and orange (Fe₂). The blue doted lines represent the movement of the structure.

</figure> <figure id="web_plot8">

'Normalized squared displacement per residue for mode 7 to 12. The x-axis represent the residue number in the sequence, whereby 0 is the first of the structure sequence, which is residue 103 in the FASTA-sequence of P00439. the y-axis shows the normalized squared displacement.

</figure>

Mode 8 of WEBnm@ show a few differences to mode 7 as there seems only be a atomic displacement at residues 20 to 50 (<xr id="web_plot8"/>). In <xr id="web_mode8"/> you can see that this is represented in the picture on the right site of the binding site. The animation looks like a hinge-movement. Maybe this helps the protein to bind or release a substrate.

Mode 9

Mode 9 captured of WEBnm@.The PAH structure (pdb 1j8u) is shown in transperent green, the ligands in red (H4B: C₉H₁₅N₅O₃) and orange (Fe₂). The blue doted lines represent the movement of the structure.

</figure> <figure id="web_plot9">

'Normalized squared displacement per residue for mode 7 to 12. The x-axis represent the residue number in the sequence, whereby 0 is the first of the structure sequence, which is residue 103 in the FASTA-sequence of P00439. the y-axis shows the normalized squared displacement.

</figure>

Like in the mode before there is only a part of the protein showing a high atomic displacement. This part is between the residues 280 and 310 (<xr id="web_plot9"/>). However, there is also a small part in the beginning which shows atomic displacement of about 2 in the normalized value. Also you can see in <xr id="web_mode9"/> that there are two parts of the protein that have a high fluctuation, which is at the top of the picture and at the right. The vectors on the right seem to show a movement inside the protein.

Mode 10

Mode 10 captured of WEBnm@. The PAH structure (pdb 1j8u) is shown in transperent green, the ligands in red (H4B: C₉H₁₅N₅O₃) and orange (Fe₂). The blue doted lines represent the movement of the structure.

</figure> <figure id="web_plot10">

'Normalized squared displacement per residue for mode 7 to 12. The x-axis represent the residue number in the sequence, whereby 0 is the first of the structure sequence, which is residue 103 in the FASTA-sequence of P00439. the y-axis shows the normalized squared displacement.

</figure>

Mode 10 shows a high atomic displacement in some parts of the protein. Especially in the beginning before residue 50 and again after residue 280 (<xr id="web_plot10"/>). However, in <xr id="web_mode10"/> there seems to be not that much movement.

Mode 11

Mode 11 captured of WEBnm@. The PAH structure (pdb 1j8u) is shown in transperent green, the ligands in red (H4B: C₉H₁₅N₅O₃) and orange (Fe₂). The blue doted lines represent the movement of the structure.

</figure> <figure id="web_plot11">

'Normalized squared displacement per residue for mode 7 to 12. The x-axis represent the residue number in the sequence, whereby 0 is the first of the structure sequence, which is residue 103 in the FASTA-sequence of P00439. the y-axis shows the normalized squared displacement.

</figure>

This mode seems to have some similarities with mode 10 in the atomic displacement as there is a high atomic displacement in the beginning and in the end of the protein (<xr id="web_plot11"/>). However, looking at <xr id="web_mode11"/> some differences can be viewed like in the bottom left of the picture, where you can see that the movement has another direction and may be indicate a small twist.

Mode 12

Mode 12 captured of WEBnm@. The PAH structure (pdb 1j8u) is shown in transperent green, the ligands in red (H4B: C₉H₁₅N₅O₃) and orange (Fe₂). The blue doted lines represent the movement of the structure.

</figure> <figure id="web_plot12">

'Normalized squared displacement per residue for mode 7 to 12. The x-axis represent the residue number in the sequence, whereby 0 is the first of the structure sequence, which is residue 103 in the FASTA-sequence of P00439. the y-axis shows the normalized squared displacement.

</figure>

The last mode in this task provided by WEBnm@ shows highest atomic displacement at residues 25 to 50 (<xr id="web_plot12"/>). In <xr id="web_mode12"/> you can see that there is movement in the whole protein.

Correlation matrix of the protein PAH (pdb 1J8U) for the residues 103 to 427 of the FASTA sequence. Red color stands for positive correlations and blue for negative.

</figure> <xr id="correlation"/> represents the correlation of the residues of the protein. You can see in the fluctuations that the whole structures moves as one indictaing that there is only one domain. This coincide with the fact that PAH consists of two domains, where the second one starts at residue 119 and ends at residue 450. The first domain is not included in the different pdb structures of PAH as it goes from residue 35 to 100.

elNémo

ElNémo computes models for the given protein 1J8U. Thereby it calculates 100 lowest-frequency modes and provides a lot of information about the modes with different parameters and visualizations:

PDB-files with the conformations of the modes
animations of the modes of three different sites
CA-vari (calculates distance fluctuations between all C-alpha atoms)
R² residue mean square displacement of all C-alpha atoms
Frequency and collectivity of the modes

First five modes
Mode	Frequency	Collectivity
mode 7	1.00	0.5475
mode 8	1.02	0.4062
mode 9	1.36	0.2941
mode 10	1.43	0.0948
mode 11	1.60	0.3466

...

</figtable>

B-factor analysis for the correlation between observed and normal-mode-derived atomi displacement parameters:
Correlation = 0.534 for 307 C-alpha atoms
RMSD: only if two structures are used

Theoretical every number of normal mode perturbed models can be generated. However, a high number would take a lot of time. On default five normal modes will be computed by elNémo and can be increased up to 25. Additionally more normal modes can be calculated after the run. The analyses of the five modes with lowest frequency normal modes are shown below. For the calculation of the models all atoms are used.

Mode 7

Representation of the PAH (pdb 1J8U) movement calculated by elNémo.

</figure>

Matrix showing the maximal distance fluctuation of PAH (pdb 1J8U). If the distances decrease they are plotted in red, whereas increasing distances are shown in blue, but only if they are under the 10% strongest distance changes.

</figure>

Like mode 7 of WEBnm@ this mode of elNémo seems to have a pumping or "breathing" movement (<xr id="mode7_gif"/>). In principal the whole protein seems to be part of the fluctuation including the β-strand shown in red. The matrix in <xr id="mode7_matrix"/> also shows both increasing and decreasing distances spread over the whole protein.

Mode 8

Representation of the PAH (pdb 1J8U) movement calculated by elNémo.

</figure> <figure id="mode8_matrix">

Matrix showing the maximal distance fluctuation of PAH (pdb 1J8U). If the distances decrease they are plotted in red, whereas increasing distances are shown in blue, but only if they are under the 10% strongest distance changes.

</figure>

In <xr id="mode8_gif"/> you can see that the protein opens and closes at the binding site, whereby it looks like an opening and closing mouth. So this mode shows a hinge-movement for our protein PAH. The β-strand (red) does not move. Additionally in the correlation matrix (<xr id="mode8_matrix"/>) only small parts of the protein seem to move at all and the distances hardly ever increase, but decrease. In WEBnm@ a similar movement can be seen in mode 8 with just a few residues moving.

Mode 9

Representation of the PAH (pdb 1J8U) movement calculated by elNémo.

</figure> <figure id="mode9_matrix">

Matrix showing the maximal distance fluctuation of PAH (pdb 1J8U). If the distances decrease they are plotted in red, whereas increasing distances are shown in blue, but only if they are under the 10% strongest distance changes.

</figure>

In this mode first you can see that the β-strand (red) moves a lot (<xr id="mode9_gif"/>). Like in mode 7 it looks like "breathing", just in a stronger way. In the matrix (<xr id="mode9_matrix"/>) you can see that the residues in the beginning seem to get higher distances to the other residues, whereas from residue 70 a smaller distance to the last 20 residues can be viewed. So with this two parts it has some similarities with mode 9 of WEBnm@.

Mode 10

Representation of the PAH (pdb 1J8U) movement calculated by elNémo.

</figure> <figure id="mode10_matrix">

Matrix showing the maximal distance fluctuation of PAH (pdb 1J8U). If the distances decrease they are plotted in red, whereas increasing distances are shown in blue, but only if they are under the 10% strongest distance changes.

</figure>

Mode 10 (<xr id="mode10_gif"/>) has some similarities with mode 8 showing a hinge-movement. Also the β-strand on the right shows no fluctuation. <xr id="mode10_matrix"/> shows some movements on different parts of the protein. In comparison with the modes found with WEBnm@ this mode has some resemblances with both mode 10 and 11.

Mode 11

Representation of the PAH (pdb 1J8U) movement calculated by elNémo.

</figure> <figure id="mode11_matrix">

Matrix showing the maximal distance fluctuation of PAH (pdb 1J8U). If the distances decrease they are plotted in red, whereas increasing distances are shown in blue, but only if they are under the 10% strongest distance changes.

</figure>

The fluctuation of mode 11 of PAH (<xr id="mode11_gif"/>) looks very similar to mode 9 (<xr id="mode9_gif"/>) with a "breathing" movement. Additionally the matrices have some resemblances. However, in this case (xr id="mode11_matrix"/>) there are more increasing distances and only at residue 180 the decreasing distance to the last 20 residues can be seen. This one seems to have no corresponding mode in WEBnm@.

References

Normal mode analysis (Phenylketonuria)

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Summary