Fig. 1: General overview of the PHARAOH computational tool.

PHARAOH was designed to assess and identify correlations between amino acid (AA) changes and a longer lifespan. a The tool uses three types of data sets, mouse/human acetylomes (left box), a collection of mammalian orthologue proteins, and the phylogenetic tree of each species based on its orthologs set (middle box), and mammalian maximal lifespans (right box). b The tool analyses changes in amino acids at each acetylation site using pairwise sequence alignment between the acetylated peptide and the orthologous protein of each mammal (upper panel), generating a replacement matrix (middle panel). Subsequently, statistical analysis and validation against the phylogenetic tree are employed to examine the correlation between amino acid replacements and a longer lifespan (lower panel). c An example of a significant acetylation site on Bphl K188 from K-to-R analysis - the central portion of the figure showcases the OrthoFinder phylogenetic tree. Surrounding the tree, the inner circle represents the lifespan of each mammal, while the outer circle displays the amino acid found at that site in each mammal. Lysine (K) in pink arginine (R) in purple, other AA in turquoise, and in green sites with no ortholog found. Color coding is shown on the right. Created in BioRender. Cohen (2025) https://BioRender.com/y84l114.