The aim of this study was to examine the effect of four commercial proteins (pea, rice, egg white and whey) on the characteristics of gluten-free layer cakes. Rice flour was partially substituted with 15, 30 and 45% protein. Hydration properties, batter density and viscosity, cake characteristics (weight loss, specific volume, texture and colour) and consumer acceptability were analysed. In general, the addition of protein increased the viscosity of the batters, with higher protein contents exhibiting greater effects and with pea protein presenting the highest effect overall. The addition of egg white protein led to the hardest cakes (p < 0.05) and whey protein, which also increased the cake hardness (p < 0.05), gave rise to cakes with the highest specific volume. Both animal proteins increased the cake cohesiveness and springiness (p < 0.05). On the contrary, pea and rice protein hardly modified hardness, colour and specific volume of cakes overall, but reduced their cohesiveness (p < 0.05). Regarding sensory evaluation, all protein-enriched cakes presented lower acceptability with respect to control cake (p < 0.05), but this effect was more pronounced when rice and egg white protein were added due to their taste, odour and texture. Whey protein cakes were, among the enriched samples, the ones with the highest acceptability.