The physicochemical properties of the emulsions prepared with four different proteins (pea protein isolate, whey protein isolate, soy protein concentrate, and soy protein isolate) were characterized in terms of particle size, rheological property, and freeze-thaw stability, while the feasibility of the protein-stabilized emulsions as a fat replacer was evaluated. Confocal laser scanning microscopic analysis showed that O/W emulsions were successfully prepared with all the proteins; however, the smallest particle size and excellent syneresis were observed in the WPI-stabilized emulsions. The use of WPI-stabilized emulsions as an alternative for shortening up to 50% level did not lead to differences in the muffin batters’ specific gravity and viscoelastic properties. After baking, the substitution of shortening with the WPI emulsions at up to 50% by weight did not significantly affect the muffins’ specific volume and texture characteristics.