The potential for using shell proteins in gastropod systematics, assessed in patellogastropod limpets

Abstract

Abstract This investigation of the application of shell protein information to gastropod systematics initially utilized available Lottia gigantea sequences and a transcriptome of Patelloida mimula developed here. Levels of differentiation between predicted sequences of reciprocal best-hit potential homologues in P. mimula and L. gigantea suggested that they could be useful within families, and possibly in higher taxa using some shell-associated proteins, particularly the peroxidases. Subsequently, proteomic analyses of the acid-soluble fraction of extractions from 17 shells and five tissue samples were conducted by combined liquid chromatography/mass spectrometry with nano-electrospray ionization. All proteins with abundance more than 1.2% in the L. gigantea shell proteome were identified with 100% confidence in most extractions by SearchGui/PeptideShaker analyses. In total, 259 of 379 peptides predicted from in silico digestion of L. gigantea shell proteins were represented by validated peptide spectrum matches in one or more specimens. Systematics applications were investigated by analysing metrics such as protein coverage by peptides and phylogenetic analyses of peptide presence/absence. The investigation suggested that diagnostic profiles based on fixed presence/absence differences can be used to separate species pairs. However, further development of analytical techniques and accumulation of reference databases is required for realising fully the systematics potential of the shell proteome.