(A) Violin plot showing the distribution (y-axis) and probability density (x-axis) of modelled myoglobin net surface charge, ZMb, among living species (black dots) of the four prevalent eulipotyphlan ecomorphotypes. (B) Correlation between ZMb and electrophoretic mobility of native myoglobin from five eulipotyphlan insectivores; data from the grey seal (Halichoerus grypus) is added for comparison. ZMb was calculated as the sum of the charge of all ionisable groups at pH 6.5 by modelling myoglobin primary structures onto the tertiary structure and using published, conserved, site-specific ionisation constants (McLellan, 1984; Mirceta et al., 2013). Electrophoretic mobility was assessed relative to the mobility of grey seal myoglobin using native polyacrylamide gel electrophoresis of heart or skeletal muscle protein extracts of the indicated species. Green, orange, brown, and blue areas (A) or symbols and fonts (B) indicate terrestrial, semi-fossorial, fossorial, and semi-aquatic/aquatic species, respectively. Phylogenetic Generalised Least Squares analysis in panel (B) revealed a highly significant positive correlation (R2 = 0.897, p<0.005) between the two parameters (solid line, y = 0.1488 x+0.3075).