(176) Assay Card

General Information
Summary Cytotoxicity of the hyen D variants.
Condition Cytotoxicity against HeLa cells was evaluated. Refer "Cytotoxicity evaluation of I11L variants" described in the method part (Du et al., 2022). Cytotoxicity was evaluated according to the cyclotide concentration required to kill half of the cells (CC50, ?M)
Result The largest changes in cytotoxic activity were for loop 2 mutants of [I11L]hyen D, followed by mutants in loop 3 and 5. CC50 of peptides on HeLa cells: hyen D: 0.92 ± 0.07; [I11L]hyen D: 1.08 ± 0.04; [G1A, I11L]hyen D: 0.69 ± 0.02; [F2A, I11L]hyen D: 2.85 ± 0.10; [G5A, I11L]hyen D: 1.29 ± 0.06; [S7A, I11L]hyen D: 4.15 ± 2.88; [V9A, I11L]hyen D: 6.01 ± 1.21; [Y10A, I11L]hyen D: 4.27 ± 0.34; [I11A]hyen D: 8.64 ± 1.21; [I11L, F14A]hyen D: 4.10 ± 0.39; [I11L, I18A]hyen D: 3.99 ± 0.21; [I11L, S21A]hyen D: 1.26 ± 0.08; [I11L, K23A]hyen D: 1.54 ± 0.08; [I11L, S24A]hyen D: 0.80 ± 0.03; [I11L, K25A]hyen D: 1.24 ± 0.06; [I11L, V26A]hyen D: 4.76 ± 0.70; [I11L, Y28A]hyen D: 1.83 ± 0.11; [I11L, K29A]hyen D: 1.02 ± 0.03; [I11L, N30A]hyen D: 0.67 ± 0.03; kB1: 6.42 ± 0.59.
AssayType Cancer Cell Toxicity

References
Du,Q., Huang,Y.H., Wang,C.K. and Craik,D.J. (2022) Mutagenesis of bracelet cyclotide hyen D reveals functionally and structurally critical residues for membrane binding and cytotoxicity. J Biol Chem 298:101822-0

Cross-references
Proteins Assayed [I11L]hyen D
[G1A, I11L]hyen D
[F2A, I11L]hyen D
[G5A, I11L]hyen D
[S7A, I11L]hyen D
[V9A, I11L]hyen D
[Y10A, I11L]hyen D
[I11A]hyen D
[I11L, F14A]hyen D
[I11L, I18A]hyen D
[I11L, S21A]hyen D
[I11L, K23A]hyen D
[I11L, S24A]hyen D
[I11L, K25A]hyen D
[I11L, V26A]hyen D
[I11L, Y28A]hyen D
[I11L, K29A]hyen D
[I11L, N30A]hyen D