(175) Assay Card

General Information
Summary Membrane binding affinity of the hyen D variants
Condition The surface plasmon resonance (SPR) lipid-binding experiments were carried out with a model membrane composed of 20% 1-palmitoyloleoyl-phosphatidylethanolamine (POPE) and 80% 1-palmitoyloleoyl-phosphatidylcholine (POPC). Peptide-to-lipid maximum binding value (P/L max, mol/mol) and the peptide concentration required to achieve half-maximum binding value (KD value, uM) of peptides.
Result The Ala mutations had varying effects on activity relative to the parent peptide [I11L]hyen D, with substitutions in loops 2 and 3 having the most impact. P/L max (mol/mol) and KD value (uM) of peptides: hyen D: 0.31 ± 0.00, 5.88 ± 0.07; [I11L]hyen D: 0.30 ± 0.00, 5.80 ± 0.03; [G1A, I11L]hyen D: 0.30 ± 0.01, 5.54 ± 0.19; [F2A, I11L]hyen D: 0.23 ± 0.01, 13.1 ± 0.88; [G5A, I11L]hyen D: 0.29 ± 0.00, 4.77 ± 0.16; [S7A, I11L]hyen D: 0.27 ± 0.01, 8.73 ± 0.50; [V9A, I11L]hyen D: 0.28 ± 0.01, 29.5 ± 1.29; [Y10A, I11L]hyen D: 0.12 ± 0.01, 55.7 ± 0.10; [I11A]hyen D: 0.26 ± 0.02; 21.5 ± 0.45; [I11L, F14A]hyen D: 0.28 ± 0.02, 19.4 ± 0.80; [I11L, I18A]hyen D: 0.19 ± 0.01, 14.7 ± 1.80; [I11L, S21A]hyen D: 0.32 ± 0.01, 4.20 ± 0.23; [I11L, K23A]hyen D: 0.28 ± 0.01, 0.28 ± 0.01; [I11L, S24A]hyen D: 0.31 ± 0.00, 4.58 ± 0.07; [I11L, K25A]hyen D: 0.27 ± 0.00, 4.80 ± 0.34; [I11L, V26A]hyen D: 0.25 ± 0.01, 14.75 ± 0.79; [I11L, Y28A]hyen D: 0.29 ± 0.01, 6.89 ± 0.36; [I11L, K29A]hyen D: 0.27 ± 0.00, 5.29 ± 0.32; [I11L, N30A]hyen D: 0.27 ± 0.00, 4.85 ± 0.27; kB1: 0.22 ± 0.00, 18.2 ± 0.53.
AssayType Membrane Binding Assay

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