Predicted Aligned Error

The Predicted Aligned Error (PAE) is a quantitative output produced by AlphaFold, a protein structure prediction system developed by DeepMind, and other similar programs. During training, the aligned error between two residues, i and j is calculated by aligning the predicted N, Cα, and C atoms of residue i onto the same atoms in the experimental structure in the training data, and measuring the resulting distance between the predicted position of the Cα atom of residue j and the experimental position of that atom. The network is trained to calculate a probability distribution over the aligned error for each pair of residues from which the PAE for each pair can be calculated. Thus, the PAE estimates the expected positional error for each residue in a predicted protein structure given the alignment of the predicted structure onto the experimental structure on a different residue. This measurement helps scientists assess the confidence in the relative positions and orientations of different parts of the predicted protein model.

Calculation and presentation

The AlphaFold2 and AlphaFold3 networks are trained to produce a probability distribution over the predicted aligned error in 64 bins,

p

i

j

b

{\displaystyle p_{ij}^{b}}

, such that bins 1-64 cover (0,0.5), (0.5,1.0),..., (31.0,31.5), (31.5-), where the last bin covers all distances larger than 31.5 Å. The sum over all 64 bins is 1.0:

∑

b

=

1

64

p

i

j

b

=

1

{\displaystyle \sum _{b=1}^{64}p_{ij}^{b}=1}

The PAE is calculated by multiplying each probability by the center value of each bin and summing:

P

A

E

i

j

=

∑

b

=

1

64

p

i

j

b

Δ

b

{\displaystyle PAE_{ij}=\sum _{b=1}^{64}p_{ij}^{b}\Delta _{b}}

where

Δ

b

=

(

b

−

0.5

)

/

2

{\displaystyle \Delta _{b}=(b-0.5)/2}

.

PAE is presented as a two-dimensional (2D) interactive plot where the color at coordinates (x, y) represents the predicted position error at residue x if the predicted and true structures were aligned on residue y. Lower PAE values for residue pairs from different domains suggest well-defined relative positions and orientations in the prediction, while higher PAE values indicate uncertainty in the relative positions or orientations.

Users can download the raw PAE data for all residue pairs in a custom JSON format for further analysis or visualization using a programming language such as Python. The format of the JSON file is as follows:

[

{

"predicted_aligned_error": [[0, 1, 4, 7, 9, ...], ...],

"max_predicted_aligned_error": 31.75

}

]

In the JSON file, the field predicted_aligned_error provides the PAE value for each residue pair (rounded to the nearest integer), and the field max_predicted_aligned_error gives the maximum possible PAE value, which is capped at 31.75 Å. The PAE is measured in Ångströms.