It is well established that the vast majority of proteins of all taxonomical groups and species are initiated by an AUG codon,translated into the amino acid methionine(Met).Many attempts were made to evaluate the importance of the sequences surrounding the initiation codon,mostly focusing on the RNA sequence.However,the role and importance of the amino acids following the initiating Met residue were rarely investigated,mostly in bacteria and fungi.Herein,we computationally examined the protein sequences of all major taxonomical groups represented in the Swiss-Prot database,and evaluated the preference of each group to specific amino acids at the positions directly following the initial Met.The results indicate that there is a species-specific preference for the second amino acid of the majority of protein sequences.Interestingly,the preference for a certain amino acid at the second position changes throughout evolution from lysine in prokaryotes,through serine in lower eukaryotes,to alanine in higher plants and animals.