|Mutation Codon||His -> Asp|
|Mutation Triplet||CAT -> GAT|
First of all, we explored the amino acid properties and compared them for the original and the mutated amino acid. Therefore we created the possible effect that the mutation could have on the protein.
|aromatic, positive charged, polar, hydrophilic||negative charged, small, polar, hydrophilic||On the one side, both amino acids are polar, but on the other side, His is positively charged, while Asp is negatively charged, which is an essential difference between these both amino acids. Therefore it is very likely, that this change causes big changes in the structure of the protein and the protein therefore will probably not work any longer. Furthermore, the structure of the two amino acids is very different, because of the aromatic ring of the His.|
Visualisation of the Mutation
In the next step, we created the visualization of the muation with PyMol. Therefore we created a picture for the original amino acid, for the new mutated amino acid and finally for both together in one picture whereas the mutation is white colored. The following pictures display that the mutated amino acid Aspartate looks very different to Histidine. Histidine has an aromatical ring. Contrary, Aspartate is smaller and forks at the end of the rest. Furthermore, it is also orientated in a completly different direction. This shows that the amino acids have huge structural differences which will probably cause drastical effects on protein structure and function.
|picture original aa||picture mutated aa||combined picture|
Subsitution Matrices Values
|PAM 1||Pam 250||BLOSOUM 62|
|value aa||most frequent substitution||rarest substitution||value aa||most frequent substitution||rarest substitution||value aa||most frequent substitution||rarest substitution|
|3||20 (Gln)||0 (Ile, Met)||4||7 (Gln)||2 (Ala, Cys, Gly, Ile, Leu, Met, Phe, Thr, Trp, Val)||-1||2 (Tyr)||-3 (Cys, Ile, Leu, Val)|
Conservation analysis with Multiple Alignments
As a next step we created a multiple alignment which contains the HEXA sequence and 9 other mammalian homologous sequences from uniprot. Afterwards we looked at the position of the different mutations and looked at the conservation level on this position. The regarded mutation is presented by the colored column. Here we can see, that the all other mammalians have on this Position the amino acid Histidine. Therefore, the mutation on this position is highly conserved and a mutation on there will cause probably huge structural and functional changes for the protein.
Secondary Structure Mutation Analysis
JPred: ...EEEECCCCCEEEEEECCCCCCCHHHHHHHHHHHHHHCCCEEEEEEECCCCCCCCC... PsiPred: ...EEECCCCCCCCCCCCCCCCCCCHHHHHHHHHHHHHCCCCEEEEEECCCCCCCEEC...
Comparison with the real structure:
|Substitution||Prediction||Reliability Index||Expected Accuracy|
A detailed list of all possible substitutions can be found [here]
Each entry contains the score at a particular position (row) for an amino acid substitution (column). Substitutions predicted to be intolerant are highlighted in red.
Threshold for intolerance is 0.05.
Amino acid color code: nonpolar, uncharged polar, basic, acidic.
Capital letters indicate amino acids appearing in the alignment, lower case letters result from prediction.