VopE, a type III effector protein of Vibrio cholerae, modulates host mitochondrial function. Mitochondrial entry of VopE is directly linked with an N-terminal precursor sequence known as the mitochondrial targeting sequence or MTS. MTS of VopE is constituted with 23 amino acids. Earlier studies have shown the importance of leucine residue at position 4 in VopE translocation to mitochondria. In the present study, we have identified another leucine residue at position 15 contributing to the mitochondrial uptake of VopE in the yeast model system. Substitution of leucine