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The identity matrix for the alignment was calculated using the PID3 method in the SIAS webmask (http://imed.med.ucm.es/Tools/sias.html).html
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A score for each column in the alignment is calculated using the concept of continuous sequence space introduced by Vingron and Sibbald [64] and the column scores are then summed over the full length of the alignment.
Positional entropy for position k in an alignment was calculated using the formula, where P jk is the frequency of amino acid j in alignment position k.
For each family, a multiple sequence alignment was calculated using POA [ 56].
The optimal structure element alignment is calculated using dynamic programming.
For each pairwise alignment, the E-value and percentile rank relative to all other pairwise alignments was calculated using SSEARCH [ 21].
BLAST [28] and PSI-BLAST [29] were used for local sequence database searches and alignments were calculated using ClustalW2 [30].
For clusters of proteins with at least 5 members from reference species, multiple sequence alignments were calculated using COBALT [ 29].
Alignments were calculated using Clustal Omega implemented in UniProt.
Initial alignments were calculated using T-COFFEE [ 62] with manual evaluations and edits in Jalview [ 63] for poorly aligning sequences.
Alignment of focal adhesions was calculated using imageJ software.
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