Part:BBa_K4359001

E.coli K-12 Cytolysin A (ClyA) (Y181F)

ClyA (Y181F) (BBa_K4359001) is a derivative of ClyA (BBa_K4359000) with a Y181F substitution mutation and the N-terminal 6X His tag and thrombin recognition and cleavage site removed. ClyA (Y181F) has improved safety over ClyA. This tyrosine residue is contained in the cholesterol-binding site in the beta-tongue motif of ClyA. The substitution is speculated to cause a loss in the flexibility of the beta-tongue motif and hence hinders the transition from the beta-tongue motif to the helix-turn-helix motif accompanying protomer formation. Y181F results in a near loss of the hemolytic activity of ClyA, as determined by erythrocyte lysis assays. This mutation is adapted from Kulshrestha, A. et al., Biorxiv, 2021. For further information on the biology and usage of this part, please refer to ClyA (BBa_K4359000).