Analysis of p.Pro731Leu variant, CFTR gene, CF Transmembrane conductance Regulator protein (1480 residues)
Data provided and calculated by CYSMA must be considered as predictions.
They are meant for educational purposes only and are provided with NO WARRANTY with respect to their biological reliability.
The wild-type residue P731 is highly conserved among the CFTR orthologs: 76% (38 / 50 CFTR orthologs)The variant P731L has been found among the CFTR orthologs with a high frequency: 10% (5 / 49 CFTR orthologs)
*AAPI: Alignment Average Percentage Identity
**AAPIR: Alignment Average Percentage Identity of the Region (20 residues surrounding position 731). AAPIR appears in green if it is more than 10% compared to AAPI, in red if less than 10%. Click here for more details on the alignment.
Divergencies show the amino acids which have been selected in the evolution.
If you find your variant among them with a high occurrence, there are good chances that your variant will most likely either have a small impact or no impact at all on the CFTR function.
Please note that CYSMA does not consider splicing alterations.
Refer to the Help page for more details.
CYSMA's visualizing modules for Ortholog conservation:
⬇ Download the region alignment (50 residues, Fasta format)
⬇ Download the CFTR phylogenic tree
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731
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Homo sapiens
S
I
V
Q
K
T
P
L
-
-
Q
M
N
G
I
-
E
E
D
S
-
-
D
E
P
L
E
R
R
L
S
L
V
P
D
-
S
E
Q
G
E
A
I
L
P
R
I
S
V
I
Pan troglodytes
S
I
V
Q
K
T
P
L
-
-
Q
M
N
G
I
-
E
E
D
S
-
-
D
E
P
L
E
R
R
L
S
L
V
P
D
-
S
E
Q
G
E
A
I
L
P
R
I
S
V
I
Pongo pygmaeus
S
I
V
Q
K
T
P
L
-
-
Q
M
N
G
I
-
E
E
D
S
-
-
D
E
P
F
E
R
R
V
S
L
V
P
D
-
S
E
Q
G
E
A
I
L
P
R
I
S
V
I
Gorilla gorilla
S
I
V
Q
K
T
P
L
-
-
Q
M
N
G
I
-
E
E
D
S
-
-
D
E
P
L
E
R
R
L
S
L
V
P
D
-
S
E
Q
G
E
A
I
L
P
R
I
S
V
I
Nomascus leucogenys
S
I
V
P
K
T
P
L
-
-
Q
M
N
G
I
-
E
E
D
S
-
-
D
E
P
L
E
R
R
L
S
L
V
P
D
-
S
E
Q
G
E
A
I
L
P
H
I
S
V
I
Macaca mulatta
S
I
V
Q
K
T
P
L
-
-
Q
M
N
G
I
-
E
E
D
S
-
-
D
E
P
L
E
R
R
L
S
L
V
P
D
-
S
E
Q
G
E
V
I
L
P
R
I
S
V
I
Macaca nemestrina
S
I
V
Q
K
T
P
L
-
-
Q
M
N
G
I
-
E
E
D
S
-
-
D
E
P
L
E
R
R
L
S
L
V
P
D
-
S
E
Q
G
E
V
I
L
P
R
I
S
V
I
Macaca fascicularis
S
I
V
Q
K
T
P
L
-
-
Q
M
N
G
I
-
E
E
D
S
-
-
D
E
P
L
E
R
R
L
S
L
V
P
D
-
S
E
Q
G
E
V
I
L
P
R
I
S
V
I
Papio anubis
S
I
V
Q
K
T
P
L
-
-
Q
M
N
G
I
-
E
E
D
S
-
-
D
E
P
L
E
R
R
L
S
L
V
P
D
-
S
E
Q
G
E
V
I
L
P
R
I
S
V
I
Callithrix jacchus
S
I
V
Q
K
T
P
L
-
-
Q
M
N
G
I
-
E
E
D
S
-
-
D
E
P
L
E
R
R
L
S
L
V
P
D
-
S
E
Q
G
E
A
I
L
P
R
I
S
V
I
Chlorocebus aethiops
S
I
V
Q
K
T
P
L
-
-
Q
M
N
G
I
-
E
E
D
S
-
-
D
E
P
L
E
R
R
L
S
L
V
P
D
-
S
E
Q
G
E
V
I
L
P
R
I
S
V
I
Colobus guereza
S
I
V
Q
K
T
P
L
-
-
Q
M
N
G
I
-
E
E
D
S
-
-
D
E
P
L
E
R
R
L
S
L
V
P
D
-
S
E
Q
G
E
V
I
L
P
R
I
S
V
I
Ateles geoffroyi
S
I
V
Q
K
T
P
L
-
-
Q
M
N
G
I
-
E
E
N
S
-
-
D
E
P
L
E
R
R
L
S
L
V
P
D
-
S
E
Q
G
E
A
I
L
P
R
I
S
V
I
Plecturocebus moloch
S
I
V
H
K
T
P
L
-
-
Q
M
N
G
I
-
E
E
D
S
-
-
D
E
P
S
E
R
R
L
S
L
I
P
D
-
S
E
Q
G
E
A
I
L
P
R
I
S
V
I
Saimiri boliviensis
S
I
V
Q
K
T
P
L
-
-
Q
M
N
G
I
-
E
E
D
S
-
-
D
E
P
L
E
R
R
L
S
L
I
P
D
-
S
E
Q
G
E
A
I
L
P
R
I
S
V
I
Aotus nancymaae
S
I
V
Q
K
T
P
L
-
-
Q
M
N
G
I
-
E
E
D
S
-
-
D
E
P
L
E
R
R
L
S
L
V
P
D
-
S
E
Q
G
E
A
I
L
P
R
I
S
M
I
Otolemur garnettii
S
V
V
Q
K
T
P
L
-
-
P
M
N
G
I
D
E
E
D
S
-
-
E
E
P
V
E
R
R
L
S
L
V
P
D
-
S
E
Q
G
E
A
I
L
P
R
S
N
V
F
Microcebus murinus
S
V
V
Q
K
T
P
L
-
-
Q
M
S
G
I
-
E
E
D
P
-
-
D
E
P
S
E
R
R
L
S
L
V
P
D
-
S
E
Q
G
E
A
I
L
P
R
S
N
V
I
Vicugna pacos
S
I
V
Q
K
T
P
L
-
-
Q
M
N
G
I
-
E
E
D
S
-
-
D
E
P
L
E
R
R
L
S
L
V
P
D
-
S
E
Q
G
E
A
I
L
P
R
S
N
V
I
Sus scrofa
S
I
V
Q
K
T
P
L
-
-
Q
M
N
G
F
-
E
E
D
S
-
-
G
E
P
L
E
R
R
L
S
L
V
P
D
-
S
E
H
G
E
A
I
L
P
R
S
N
V
I
Bos taurus
S
V
V
Q
K
T
S
L
-
-
Q
M
N
G
I
-
E
G
A
A
-
-
D
A
P
L
E
R
R
L
S
L
V
P
H
-
S
E
P
G
E
G
I
L
P
R
S
N
A
V
Muntiacus reevesi
S
V
A
Q
K
T
S
L
-
-
Q
M
N
G
I
-
E
E
T
S
-
-
D
E
P
L
E
R
K
L
S
L
V
P
H
-
S
E
P
G
E
G
I
L
P
R
S
N
A
V
Muntiacus muntjak
S
V
A
Q
K
T
S
L
-
-
Q
M
N
G
I
-
E
E
T
S
-
-
D
E
P
L
E
R
K
L
S
L
V
P
H
-
S
E
P
G
E
G
I
L
P
R
S
N
A
V
Ovis aries
S
V
V
Q
K
T
S
L
-
-
Q
M
N
G
I
-
D
G
A
S
-
-
D
E
P
L
E
R
R
L
S
L
V
P
H
-
S
E
P
G
E
G
I
L
P
R
S
N
A
V
Equus caballus
S
I
V
Q
K
T
P
L
-
-
Q
M
N
G
I
-
E
E
D
S
-
-
D
E
P
L
E
R
R
L
S
L
V
P
D
-
S
E
Q
G
E
A
I
L
P
R
S
N
V
I
Canis familiaris
S
I
V
Q
K
T
P
L
-
-
P
M
N
G
I
-
E
G
E
S
-
-
E
V
P
V
E
R
R
L
S
L
F
P
D
-
C
E
Q
G
E
A
I
L
P
R
S
N
M
I
Loxodonta africana
S
V
V
Q
K
T
P
L
-
-
Q
M
N
G
I
-
D
E
D
S
-
-
D
E
P
L
E
R
R
L
S
L
V
P
D
-
S
E
Q
G
E
A
I
L
P
R
S
N
V
I
Mustela furo
S
V
V
Q
K
T
P
L
-
-
Q
M
N
G
I
-
E
E
D
S
-
-
D
E
P
L
E
R
R
L
S
L
V
P
D
G
A
E
Q
G
E
A
I
L
P
R
S
N
M
I
Oryctolagus cuniculus
S
I
V
P
K
T
P
L
-
-
Q
M
N
G
I
-
E
E
D
S
-
-
D
A
S
I
E
R
R
L
S
L
V
P
D
-
S
E
Q
G
E
A
I
L
P
R
S
N
M
I
Atelerix albiventris
S
I
V
Q
K
T
T
L
-
-
Q
M
N
G
I
-
E
A
N
S
-
-
E
E
P
L
E
R
R
L
S
L
V
P
D
-
S
E
L
G
E
T
I
L
P
R
S
N
V
I
Dasypus novemcinctus
S
I
T
Q
K
T
P
L
-
-
P
M
N
G
I
-
D
E
D
S
-
-
G
E
T
L
E
R
R
L
S
L
V
P
D
-
C
E
Q
G
E
G
I
L
P
R
S
N
L
I
Rhinolophus ferrumequinum
S
I
V
Q
K
T
P
L
-
-
Q
M
N
G
I
-
E
E
D
S
-
-
E
E
S
S
E
R
R
L
S
L
V
P
D
-
S
E
Q
G
E
T
I
L
P
R
S
N
L
I
Cavia porcellus
S
I
V
P
K
T
P
L
-
-
Q
I
S
G
I
-
E
E
D
S
-
-
D
D
P
V
E
R
R
L
S
L
V
P
D
-
S
E
Q
S
D
G
L
L
L
R
S
N
V
I
Monodelphis domestica
S
V
V
Q
K
S
Q
P
-
-
Q
M
N
G
I
-
E
E
N
D
-
-
D
E
P
L
E
R
R
L
S
L
V
P
D
-
S
E
Q
G
E
A
I
L
P
R
S
N
I
M
Ornithorhynchus anatinus
S
L
V
Q
K
A
P
L
-
-
Q
M
N
C
I
-
D
E
T
L
-
-
D
E
P
P
E
R
K
L
S
L
V
P
D
-
S
E
P
G
E
A
I
L
P
R
S
N
M
L
Didelphis virginiana
S
V
V
Q
K
G
Q
P
-
-
Q
M
N
G
I
-
E
E
N
D
-
-
D
E
P
L
E
R
R
L
S
L
V
P
D
-
S
E
Q
G
E
A
I
L
P
R
S
N
I
I
Trichosurus vulpecula
S
V
V
Q
K
G
Q
Q
-
-
Q
M
N
G
I
-
E
E
N
D
-
-
D
E
P
L
E
R
R
L
S
L
V
P
D
-
S
E
Q
G
E
A
I
L
P
R
S
N
M
I
Carollia perspicillata
S
V
V
Q
K
T
P
L
-
-
Q
M
N
G
I
-
E
E
D
S
-
-
E
E
P
S
E
R
R
L
S
L
V
P
D
-
S
E
Q
G
E
A
I
L
P
R
S
N
V
I
Mus musculus
S
I
V
Q
K
T
P
L
-
-
C
I
-
-
-
-
D
G
E
S
-
-
D
D
L
Q
E
K
R
L
S
L
V
P
D
-
S
E
Q
G
E
A
A
L
P
R
S
N
M
I
Rattus norvegicus
S
I
V
Q
K
T
P
L
-
-
S
I
-
-
-
-
E
G
E
S
-
-
D
D
L
Q
E
R
R
L
S
L
V
P
D
-
S
E
H
G
E
A
A
L
P
R
S
N
M
I
Gallus gallus
S
I
M
Q
K
N
G
T
-
-
Q
V
N
G
L
-
E
D
G
H
-
-
I
D
S
P
E
R
R
I
S
L
V
P
D
-
L
E
Q
G
D
V
G
L
P
R
S
N
M
L
Taeniopygia guttata
S
V
V
Q
K
N
G
M
-
-
Q
V
N
G
L
-
E
D
G
H
-
-
S
D
P
A
E
R
R
L
S
L
V
P
D
-
V
E
Q
G
D
V
A
L
L
H
S
G
M
L
Xenopus tropicalis
S
I
M
Q
K
S
Q
P
-
-
Q
M
S
G
I
-
E
E
E
D
-
-
I
P
P
N
E
R
K
L
S
L
V
P
E
-
S
E
Q
G
E
A
S
L
P
R
S
N
V
L
Xenopus laevis
S
L
M
Q
K
S
Q
P
-
-
Q
M
S
G
I
-
E
E
E
D
M
P
A
E
Q
G
E
R
K
L
S
L
V
P
E
-
S
E
Q
G
E
A
S
L
P
R
S
N
F
L
Squalus acanthias
S
L
V
Q
T
A
M
S
Y
P
Q
T
N
G
M
-
E
D
A
T
-
-
S
E
P
G
E
R
H
F
S
L
I
P
E
-
N
E
L
G
E
P
T
K
P
R
S
N
I
F
Danio rerio
-
-
-
-
K
A
S
F
-
-
I
Q
V
P
E
-
E
E
V
R
-
-
R
T
L
P
D
R
K
F
S
L
V
P
E
-
N
E
L
V
D
E
S
F
M
G
S
D
V
Y
Oryzias latipes
S
F
I
G
T
G
T
L
-
-
Q
P
S
L
M
-
E
E
G
G
-
-
N
E
L
S
D
R
R
F
S
V
V
P
E
-
D
D
Q
V
E
E
A
L
P
R
S
N
L
Y
Takifugu rubripes
-
-
-
-
S
Q
A
A
-
-
Q
A
T
A
T
-
E
D
G
V
-
-
H
E
H
S
E
R
R
F
S
V
V
P
E
-
D
D
Q
V
E
E
V
L
P
R
S
N
A
Y
Tetraodon nigroviridis
S
F
M
G
G
N
A
T
-
-
Q
S
M
A
P
-
E
E
G
V
-
-
H
E
L
S
E
R
R
F
S
V
V
P
E
-
D
D
Q
V
E
E
V
L
P
R
S
N
V
Y
Caenorhabditis elegans
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
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Species color legend (basic classification):
Great apes | Other monkeys | Prosimians | Other mammals | Lizards | Birds | Amphibians | Fishes | Insects | Nematods | Tunicates | Echinoderms
Ortholog sequences have been selected from the Ensembl(1) and
NCBI websites. Alignment has been performed with
ClustalW(2), version 1.83 or 2.0.7.
Trees have been built using Phylogeny.fr(3), based on the alignments.
Software used is PhyML 3.0 aLRT with default parameters. Pictures of trees have been made using Phylip at Mobyle.
AAPI and AAPIR have been calculated thanks to Bioperl.
Domain conservation:
The residue p.Pro731 belongs to the domain R.
650
842
the regulatory (R) domain links the two transporter halves (MSD1-NBD1 and MSD2-NBD2). It is a highly flexible region and it contains multiple phosphorylation sites, which are the target of Protein Kinase A and B (PKA and PKC), and control the gating and traffiking of the chloride chanel.
R: the regulatory (R) domain links the two transporter halves (MSD1-NBD1 and MSD2-NBD2). It is a highly flexible region and it contains multiple phosphorylation sites, which are the target of Protein Kinase A and B (PKA and PKC), and control the gating and traffiking of the chloride chanel.
R of CF Transmembrane conductance Regulator domain alignment including p.Pro731 residue.
***AAPID: Alignment Average Percentage Identity of the Domain (positions are indicated). !AAPIR: Alignment Average Percentage Identity of the Region (20 residues surrounding position 731). AAPIR appears in green if it is more than 10% compared to AAPID, in red if less than 10%.
Divergencies
Residues present in more than 10% of the sequences are highlighted in blue.
H - 2.13%
L - 10.64%
Q - 2.13%
S - 6.38%
T - 2.13%
The wild-type residue P731 belongs to the R domain and is conserved at 76.60% among the R homologs (36 / 47 R homologs)
The variant P731L has been found among the R homologs with a high frequency: 10.64% (5 / 47 R homologs)
Divergencies show the amino acids which have been selected in the evolution. Residues present in more than 10% of the sequences are highlighted in blue.
Please note that CYSMA does not consider splicing alterations.
Refer to the Help page for more details.
CYSMA's visualizing modules for R domain conservation:
Sequence alignments for NBDs have all been extracted from Prosite(4). Sequence alignments for MSDs have been extracted using the PSI-BLAST web server.
Sequences alignments have been manually re-aligned using a structural alignment including the human CFTR and bacterian ABC transporters with know 3D structures (for MSDs and NBDs domains).
Predictions of secondary structures have been made with PsiPred(9)
, version 2.5, using Protein Multiple Sequences Alignments as input, in order to increase the accuracy of the prediction.
Amino acid frequencies have been calculated from a non redondant set defined by the RCSB.
The Help page will tell you more about it.
3D analysis:
Models provided and analysed by CYSMA must be considered as predictions, therefore be careful when interpreting the results. All efforts have been made to build structures of quality, however, they are provided with NO WARRANTY as to their accuracy with the real biological molecules studied.
Wild type and predicted mutant structures have been compared. You will find the results below.
Click on the MolProbity logo for complete details on the structure quality
This model is made of 37 α helices and of 20 β strands (and is mainly composed of helices (739 residues in helices against 102 in strands, for a total of 1480 amino acids)).
3D structures predicts P731 to be located in a loop (which confirms PsiPred prediction) and L731 in a loop.
WARNING! The experimental 3D structure used for our predictions is the complete human CFTR structure which have been solved at a 3.7 Å resolution using cryo-electron microscopy (PDB: 5UAK; Liu et al. 2017). The overall resolution is fairly low so the CYSMA's 3D Automatic Annotation pipeline might have missed some important structural effects.
Replacement of a proline is likely to increase the flexibility of the region
Solvent accessibility: the wild-type P731 and the mutant P731L are predicted to be buried
Hydrogen bond network:
P731
L731
none
none
The mutant residue might introduce steric clashes
P731
L731
none
2.84 Å between LEU 731 CB and LEU 732 N
CYSMA's 3D visualizing module:
If you want to investigate further the structures, you can use
the JSmol applets of the wild-type (left) and mutant (right) structures.
Click on the JSmol applets' link to hide it.
You have a full access to Jmol commands with a simple right click on one applet.
JSmol Legends:
The residue at the position 731 is located in the center, labelled in yellow and surrounded by its neighboring residues (distance < 5 Å).
Van der Waals contacts with the residue 731 are represented by dotted lines.
Amino acids involved in H-bonds with the residue 731 are labelled in blue.
Amino acids involved in steric clashes with the residue 731 are labelled in red.
The overall structure of the complete human CFTR is represented in ribbon diagrams (click on the Reset button to visualize the overall CFTR structure). The membrane-spanning domain MSD1 is represented in blue and MSD2 in light blue.The nucleotide-binding domain NBD1 is represented in orange, NBD2 in light salmon.The lasso domain is shown in red and the R domain in green.
The 3D structures used in CYSMA are models based on the CFTR experimental 3D structure in the channel-closed conformation (PDB: 5UAK; resolution: 3.9 Å). In the wild-type model, the (missing) loops and the (missing) R domain were built de novo using the software Modeller. For the variant models, the point mutation (homology modelling) are made on the fly with Modeller (more).
Each structure has been assessed with MolProbity(19).
Msms(20) is used to calculate solvent accessibility, and STRIDE(21) (plus stride2pdb)
for secondary structure assignment.
Secondary structure analyses in 3D models uses side chain interaction energies reviewed in (23), as well as amino-acids propensities for N-caps, N1-N3, helix middle, C3-C1 and C-caps extracted from (24)(PDB values).
Structural properties are calculated using an in-house developped program based for the USMA's 3D Automatic Annotation pipeline.
Click on the LOVD picture to check if a variant is described at position 731
Graphical display of the region at NCBI (including SNPs)
CYSMA Report:
Report for p.Pro731Leu variant
CFTR orthologs conservation
The wild-type residue P731 is highly conserved among the CFTR orthologs: 76% (38 / 50 CFTR orthologs)The variant P731L has been found among the CFTR orthologs with a high frequency: 10% (5 / 49 CFTR orthologs)
R homologs conservation
The wild-type residue P731 belongs to the R domain and is conserved at 76.60% among the R homologs (36 / 47 R homologs)
The variant P731L has been found among the R homologs with a high frequency: 10.64% (5 / 47 R homologs)
Structural effects
Replacement of a proline is likely to increase the flexibility of the region
Solvent accessibility: the wild-type P731 and the mutant P731L are predicted to be buried
The mutant residue might introduce steric clashes
Allele frequency
The variant P731L in gnomAD (123,136 exomes): 5.17e-05
Clinical significance
The variant P731L has not been reported in ClinVar
CFTR-France
The variant P731L might correspond to: NM_000492.3:c.2192C>T, which is reported to be unclassified (CFTR-France for more details)
Additional resources
SIFT prediction: variant P731L is predicted to be tolerated (score: 1)
PPH2 prediction: variant P731L is predicted to be benign (score: 0)
