Sequence-based mutation analysis HEXA

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Revision as of 17:36, 23 June 2011 by Uskat (talk | contribs) (rs121907967: Trp -> TER)

Mutations

SNP-id codon number mutation codon mutation triplet
rs4777505 29 Asn -> Ser AAC -> AGC
rs121907979 39 Leu -> Arg CTT -> CGT
rs61731240 179 His -> Asp CAT -> GAT
rs121907974 211 Phe -> Ser TTC -> TCC
rs61747114 248 Leu -> Phe CTT -> TTT
rs1054374 293 Ser -> Ile AGT -> ATT
rs121907967 329 Trp -> TER TGG -> TAG
rs1800430 399 Asn -> Asp AAC -> GAC
rs121907982 436 Ile -> Val ATA -> GTA
rs121907968 485 Trp -> Arg gTGG -> CGG

Analysis of the mutations

We created for each mutation an extra page. The summary of the analysis can be seen in the Summary Section.

[rs4777505: Asn -> Ser]

[rs121907979: Leu -> Arg]

[rs61731240: His -> Asp]

[rs121907974: Phe -> Ser]

[rs61747114: Leu -> Phe]

[rs1054374: Ser -> Ile]

[rs121907967: Trp -> TER]

[rs1800430: Asn -> Asp]

[rs121907982: Ile -> Val]

[rs121907968: Trp -> Arg]


rs4777505: Asn -> Ser

pysicochemical properities

Asn Ser consequences
polar, small, hydrophilic, negatively charged polar, tiny, hydrophilic, neutral Both amino acids are polar and hydrophilic. Ser is tiny, Asn therefore is a small amino acid. The biggest difference between these two amino acid is, that Asn is negatively charged and Ser is neutral. But this is not that big difference and therefore we suggest, that this mutation do not delete the structure and function of the protein.

Visualisation of the mutation

picture original aa picture mutated aa combined picture
Amino acid Asparagine
Amino acid Serine
Picture which visualize the mutation

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
20 36 (Asp) 0 (Cys, Met) 5 7 (Asp) 2 (Cys, Leu, Phe, Trp) 1 1 (Asp, His, Ser) -4 (Trp)

Conservation analysis with multiple alignments

Mutation in the multiple alignment

Secondary structure mutation analysis

JPred:
CCHHHHHHHHHHHHHCCCCCCCEEEEEEEEEECCCEEEEECCCEEEEECCCCCCC...
PsiPred:
CHHHHHHHHHHHHHHHCCCCCCCCCCCCEEEECCCEEEEECCCEEEEECCCCCCC...

Comparison with the real structure:

Mutation at position 29
Mutation at position 29 - detailed view


SNAP Prediction

Substitution Prediction Reliability Index Expected Accuracy
S Neutral 8 96%

A detailed list of all possible substitutions can be found [here]

SIFT Prediction

SIFT Matrix:
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.

Sift legend.png
29 sift.png.png

SIFT Table
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.



Predict Not ToleratedPositionSeq RepPredict Tolerated
w29N0.78cfMHiYpVgLRNdQTASKE



Polyphen2 Prediction

HumDiv prediction
HumVar prediction

rs121907979: Leu -> Arg

Pysicochemical Properities

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.

Leu Arg consequences
aliphatic, hydrophobic, neutral positive charged, polar, hydrophilic Leucine is smaller and without a positive charge. Therefore, Arg is too big for the position of Leu, therefore, the change of Leu with Arg has to cause changes in the 3D structure of the protein. Furthermore, Leu is a hydrophobic amino acid, whereas Arg is hydrophilic. This is the complete contrary and therefore we suggest, that the protein will not function any longer.

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 Arginine has a longer chain than Leucine. Contrary Leucine comes to a fork at the end of its rest. This shows that the amino acids have some structural differences that are not drastical, but can be essential.

picture original aa picture mutated aa combined picture
Amino acid Leucine
Amino acid Arginine
Picture which visualize the mutation

Subsitution Matrices values

Afterwards, we looked at the values of the substitution matrices PAM1, PAM250 and BLOSSUM62. Therefore we looked detailed at the three values: the value for accoding amino acid substitution, the most frequent value for the substitution of the examined amino acid and the rarest substitution.

In this case, the substitution of Leucine to Arginine has very low values that are near the values for the rarest subsitution for PAM1 and PAM250. Furthermore, the value for the most frequent substitution differs also a lot from the value for this certain mutation for both PAMs. Contrary for BLOSUM62 the value for the amino acid subsitution Leucine to Arginine is average. This means the most frequent subsitution value is as far as the rarerest subsitution from the the underlying value. The difference between the two PAMs and BLOSUM62 can be ascribed to the different preparations of these two kind of substitutions matrices. The PAM-matrices is an evolutionary model whereas BLOSUM is based on protein famalies. Therefore probably this mutation is evolutionary not that unlikely whereas within a protein family it is unusual.

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
1 22 (Ile) 0 (Asp, Cys) 4 20 (Met) 2 (Cys) -2 0 (Phe) -4 (Asp, Gly)


Conservation analysis with multiple alignments

Mutation in the multiple alignment

Secondary structure mutation analysis

JPred:
CCHHHHHHHHHHHHHCCCCCCCEEEEEEEEEECCCEEEEECCCEEEEECCCCCCC...
PsiPred:
CHHHHHHHHHHHHHHHCCCCCCCCCCCCEEEECCCEEEEECCCEEEEECCCCCCCC...

Comparison with the real structure:

Mutation at position 39
Mutation at position 39 - detailed view


SNAP Prediction

Substitution Prediction Reliability Index Expected Accuracy
R Non-neutral 5 87%

A detailed list of all possible substitutions can be found [here]


SIFT Prediction

SIFT Matrix:
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.

Sift legend.png
39 sift.png.png

SIFT Table
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.



Predict Not ToleratedPositionSeq RepPredict Tolerated
hdwqpnecrgskytafm39L0.79VIL



Polyphen2 Prediction

HumDiv prediction
HumVar prediction

rs61731240: His -> Asp

pysicochemical properities

His Asp consequences
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

picture original aa picture mutated aa combined picture
Amino acid Histidine
Amino acid Aspartate
Picture which visualize the mutation


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

Mutation in the multiple alignment

Secondary structure mutation analysis

JPred:
...EEEECCCCCEEEEEECCCCCCCHHHHHHHHHHHHHHCCCEEEEEEECCCCCCCCC...
PsiPred:
...EEECCCCCCCCCCCCCCCCCCCHHHHHHHHHHHHHCCCCEEEEEECCCCCCCEEC...

Comparison with the real structure:

Mutation at position 179
Mutation at position 179 - detailed view

SNAP Prediction

Substitution Prediction Reliability Index Expected Accuracy
D Non-neutral 6 93%

A detailed list of all possible substitutions can be found [here]

SIFT Prediction

SIFT Matrix:
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.

Sift legend.png
179 sift.png.png

SIFT Table
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.



Predict Not ToleratedPositionSeq RepPredict Tolerated
mwfciyvltasperndkgQ179H0.99H



Polyphen2 Prediction

HumDiv prediction
HumVar prediction

rs121907974: Phe -> Ser

pysicochemical properities

Phe Ser consequences
polar, tiny, hydrophilic, neutral aliphatic, hydrophobic, neutral Ile is much bigger than Ser and also is branched, because it is an aliphatic amino acid. Therefore the structure of both amino acids is really different and Ile is to big for the position where Ser was. Therefore, there has to be a big change in the 3D structure of the protein and the protein probably will loose its function.

Visualisation of the mutation

picture original aa picture mutated aa combined picture
Amino acid Phenylalanine
Amino acid Serine
Picture which visualize the mutation

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
2 28 (Tyr) 0 (Asp, Cys, Glu, Lys, Pro, Val) 2 20 (Tyr) 1 (Arg, Asp, Cys, Gln, Glu, Gly, Lys, Pro) -2 3 (Tyr) -4 (Pro)

Conservation analysis with multiple alignments

Mutation in the multiple alignment


Secondary structure mutation analysis

JPred:
...EEEECCCCCEEEEEECCCCCCCHHHHHHHHHHHHHHCCCEEEEEEECCCCCCCCC...
PsiPred:
...EEECCCCCCCCCCCCCCCCCCCHHHHHHHHHHHHHCCCCEEEEEECCCCCCCEEC...

Comparison with the real structure:

Mutation at position 211
Mutation at position 211 - detailed view

SNAP Prediction

Substitution Prediction Reliability Index Expected Accuracy
S Non-neutral 5 87%

A detailed list of all possible substitutions can be found [here]


SIFT Prediction

SIFT Matrix:
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.

Sift legend.png
211 sift.png.png

SIFT Table
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.



Predict Not ToleratedPositionSeq RepPredict Tolerated
ywvtsrqpnmlkihgedca211F1.00F



Polyphen2 Prediction

HumDiv prediction
HumVar prediction

rs61747114: Leu -> Phe

pysicochemical properities

Leu Phe consequences
aliphatic, hydrophobic, neutral aromatic, hydrophobic, neutral Leu is an aliphatic amino acid, wheras Phe is an aromatic amino acid. This means, that Phe has an aromatic ring in its structure. But both amino acids are relatively big and so it is possible, that the exchange of this amino acids do not change the structure of the protein that much. Therefore, we suggest it is possible, that the protein will work.

Visualisation of the mutation

picture original aa picture mutated aa combined picture
Amino acid Leucine
Amino acid Phenylalanine
Picture which visualize the mutation

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
13 45 (Met) 0 (Asp, Cys) 13 20 (Met) 2 (Cys) 0 2 (Ile, Met) -4 (Asp, Gly)

Conservation analysis with multiple alignments

Mutation in the multiple alignment


Secondary structure mutation analysis

JPred:
...HHHHHHHHCCCEEEECCCCCHHHHHHHHHCCCCCCCCCCCCCCCCCCCCCCCCCC...
PsiPred:
...HHHHHHHHCCCEEEECCCCCHHHHHHHHCCCCCCCCCCCCCCCCCCCCCCCCCCH...

Comparison with the real structure:

Mutation at position 248
Mutation at position 248 - detailed view


SNAP Prediction

Substitution Prediction Reliability Index Expected Accuracy
F Neutral 3 78%

A detailed list of all possible substitutions can be found [here]

SIFT Prediction

SIFT Matrix:
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.

Sift legend.png
248 sift.png.png

SIFT Table
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.



Predict Not ToleratedPositionSeq RepPredict Tolerated
cwpenkgrsthaQ248L1.00YIDVFML




Polyphen2 Prediction

HumDiv prediction
HumVar prediction

rs1054374: Ser -> Ile

pysicochemical properities

Ser Ile consequences
polar, tiny, hydrophilic, neutral aliphatic, hydrophobic, neutra Ile is much bigger than Ser and also is branched, because it is an aliphatic amino acid. Therefore the structure of both amino acids is really different and Ile is to big for the position where Ser was. Therefore, there has to be a big change in the 3D structure of the protein and the protein probably will loose its function.


Visualisation of the mutation

picture original aa picture mutated aa combined picture
Amino acid Serine
Amino acid Isoleucine
Picture which visualize the mutation

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
2 38 (Thr) 1 (Leu) 5 9 (Ala, Gly, Pro, Thr) 3 (Phe) -2 1 (Ala, Asn, Thr) -3 (Trp)

Conservation analysis with multiple alignments

Mutation in the multiple alignment


Secondary structure mutation analysis

JPred:
...HHHHHHHHCCCEEEECCCCCHHHHHHHHHCCCCCCCCCCCCCCCCCCCCCCCCCC...
PsiPred:
...HHHHHHHHCCCEEEECCCCCHHHHHHHHCCCCCCCCCCCCCCCCCCCCCCCCCCH...

Comparison with the real structure:

Mutation at position 293
Mutation at position 293 - detailed view

SNAP Prediction

Substitution Prediction Reliability Index Expected Accuracy
I Neutral 2 69%

A detailed list of all possible substitutions can be found [here]

SIFT Prediction

SIFT Matrix:
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.

Sift legend.png
293 sift.png.png

SIFT Table
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.



Predict Not ToleratedPositionSeq RepPredict Tolerated
wghydrnfqekcp293S1.00MlASVTI



Polyphen2 Prediction

HumDiv prediction
HumVar prediction

rs121907967: Trp -> TER

pysicochemical properities

Trp TER consequences
aromatic, polar, hydrophobic TER By this change, the protein is not complete, therefore it is not possible for the protein to fold and to function.


Visualisation of the mutation

picture original aa picture mutated aa combined picture
Amino acid Tryptophan
Visualization of the mutated protein

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
X 2 (Arg) 0 (all, except Arg, Phe, Ser, Tyr) X 2 (Arg) 0 (all, except Arg, His, Leu, Phe, Ser, Tyr) X 2 (Tyr) -4 (Asn, Asp, Pro)

Conservation analysis with multiple alignments

Mutation in the multiple alignment


Secondary structure mutation analysis

JPred:
...CCCCCCCCCCCCHHHHHHHHHCCCCCCHHHHHHHHHHHHHHHHHHCCCEEEEECC...
PsiPred:
...CCCCCCCCCCCCHHHHHHHHHCCCCCCHHHHHHHHHHHHHHHHHHCCCCEEEECC...

Comparison with the real structure:

Mutation at position 329
Mutation at position 329 - detailed view


SNAP Prediction

No prediction available, because the protein ends here.

A detailed list of all possible substitutions can be found [here]

SIFT Prediction

SIFT Matrix:
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.

Sift legend.png
329 sift.png.png



Polyphen2 Prediction

HumDiv prediction
HumVar prediction

rs1800430: Asn -> Asp

pysicochemical properities

Asn Asp consequences
polar, small, hydrophilic, negatively charged polar, small, hydrophilic, negatively charged Both amino acids have the same properities and therefore we suggest that an exchange of these two amino acids do not destroy the protein structure and function

Visualisation of the mutation

picture original aa picture mutated aa combined picture
Amino acid Asparagine
Amino acid Aspartic acid
Picture which visualize the mutation

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
36 36 (Asp) 0 (Cys, Met) 7 7 (Asp) 2 (Cys, Leu, Phe, Trp) 1 1 (Asp, His, Ser) -4 (Trp)

Conservation analysis with multiple alignments

Mutation in the multiple alignment


Secondary structure mutation analysis

JPred:
...CCCCCCCEEEEEECCCCCCCCHHHHHHHHHCCCCEEECCCCCCCCCCCCCCCCCC...
PsiPred:
...CCCCCCCCEEEEECCCCCCCHHHHHHHHHHCCCEEEECCCCCCCCCCCCCCCCCC...

Comparison with the real structure:

Mutation at position 399
Mutation at position 399 - detailed view


SNAP Prediction

Substitution Prediction Reliability Index Expected Accuracy
D Neutral 1 60%

A detailed list of all possible substitutions can be found [here]

SIFT Prediction

SIFT Matrix:
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.

Sift legend.png
399 sift.png.png

SIFT Table
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.



Predict Not ToleratedPositionSeq RepPredict Tolerated
wyfcm399N0.72iHvlgPtRDEQASKN



Polyphen2 Prediction

HumDiv prediction
HumVar prediction

rs121907982: Ile -> Val

pysicochemical properities

Ile Val consequences
aliphatic, hydrophobic, neutra aliphatic, hydrophobic, neutral In this case, the pysicochemical properties are equal. Furthermore, they almost agree in their size. Therefore, we suggest, that there is no big effect on the 3D structure of the protein and therefore, also no big effect on the protein function.

Visualisation of the mutation

picture original aa picture mutated aa combined picture
Amino acid Isoleucine
Amino acid Valin
Picture which visualize the mutation

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
33 33 (Val) 0 (Gly, Pro, Trp) 9 9 (Val) 1 (Trp) 3 3 (Val) -4 (Gly)

Conservation analysis with multiple alignments

Mutation in the multiple alignment


Secondary structure mutation analysis

JPred:
...HHHHHHHHCCCCEEECCCCCCCCCCCCCCCCCCCCCCCCCCCCCCCCCCCEEEE...
PsiPred:
...HHHHHHHHCCCEEEECCCCCCCCCCCCCCCCCCCCCCCCCCCCCHHHHCCCCCE...

Comparison with the real structure:

Mutation at position 436
Mutation at position 436 - detailed view


SNAP Prediction

Substitution Prediction Reliability Index Expected Accuracy
V Neutral 0 53%

A detailed list of all possible substitutions can be found here]

SIFT Prediction

SIFT Matrix:
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.

Sift legend.png
436 sift.png.png

SIFT Table
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.



Predict Not ToleratedPositionSeq RepPredict Tolerated
wc436I1.00fYHMpIVgLnTQDRASEK



Polyphen2 Prediction

HumDiv prediction
HumVar prediction

rs121907968: Trp -> Arg

pysicochemical properities

Trp Arg consequences
aromatic, polar, hydrophobic, neutral positive charged, polar, hydrophilic Trp is very big, because of two aromatic rings in its structure. Furthermore, it is hydrophobic, whereas, Arg is a hydrophilic amino acid. Therefore, the changes in the 3D structure might be extreme and delete the function of the protein.

Visualisation of the mutation

picture original aa picture mutated aa combined picture
Amino acid Tryptophan
Amino acid Arginine
Picture which visualize the mutation

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
2 2 (Arg) 0 (all, except Arg, Phe, Ser, Tyr) 2 2 (Arg) 0 (all, except Arg, His, Leu, Phe, Ser, Tyr) -3 2 (Tyr) -4 (Asn, Asp, Pro)

Conservation analysis with multiple alignments

Mutation in the multiple alignment


Secondary structure mutation analysis

JPred:
...HHHHCCCCCCCCHHHHHHHHHHHHHHHHHCCCCCCCCCCCCCCHHHCCC...
PsiPred:
...HHHCCCCCCCCCHHHHHHHHHHHHHHHHHCCCCCCCCCCCCCCCCCCCC...

Comparison with the real structure:

Mutation at position 485
Mutation at position 485 - detailed view

SNAP Prediction

Substitution Prediction Reliability Index Expected Accuracy
R Non-neutral 7 96%

A detailed list of all possible substitutions can be found [here]

SIFT Prediction

SIFT Matrix:
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.

Sift legend.png
485 sift.png.png

SIFT Table
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.



Predict Not ToleratedPositionSeq RepPredict Tolerated
yvtsrqpnmlkihgfedca485W0.97W



Polyphen2 Prediction

HumDiv prediction
HumVar prediction

Summary page

method  mutations
Asn -> Ser Leu -> Arg His -> Asp Phe -> Ser Leu -> Phe Ser -> Ile Trp -> TER Asn -> Asp Ile -> Val Trp -> Arg
pysiochemical properitites neutral non-neutral non-neutral non-neutral neutral non-neutral non-neutral neutral neutral non-neutral
visuale analysis todo non-neutral non-neutral non-neutral todo neutral non-neutral todo neutral non-neutral
PAM1 neutral non-neutral non-neutral non-neutral no statement non-neutral no information neutral neutral neutral
PAM250 neutral non-neutral no statement non-neutral no statement no statement no information neutral neutral neutral
BLOSUM62 neutral no statement no statement non-neutral neutral non-neutral no information neutral neutral non-neutral
multiple alignment neutral neutral non-neutral non-neutral non-neutral neutral neutral neutral neutral non-neutral
analysis with Jpred non-neutral non-neutral neutral neutral neutral neutral neutral neutral neutral neutral
analysis with PsiPred non-neutral non-neutral neutral neutral neutral neutral neutral non-neutral neutral neutral
analysis with real structure non-neutral no statement neutral neutral non-neutral neutral non-neutral no statement non-neutral non-neutral non-neutral non-neutral
SNAP Prediction neutral non-neutral non-neutral non-neutral neutral neutral no information neutral neutral non-neutral
SIFT Prediction neutral non-neutral non-neutral non-neutral neutral neutral no information neutral neutral non-neutral
PolyPhen2 Prediction neutral non-neutral non-neutral non-neutral neutral neutral no information neutral neutral non-neutral