(106) Assay Card

General Information
Summary Substitution of certain residues of kalata B1 to alanine result in loss of insecticidal activity
Condition Insecticidal activity of the alanine mutants was assessed by measuring their impact on the development of first and second instars of Drosophila melanogaster to adulthood. This assay consumed ~3 mg per test compound by using triplicates at two peptide concentrations. Triplicates were found to vary by <10%.
Result Relative to 100% control of wild-type peptide at 230 ppm (80 uM) peptide (LD50 = 150 ppm = 52 uM). kB1 (100%), G1A (102%), L2A (100%), P3A (31%), V4A (115%), G6A (2%), E7A (0%), T8A (15%), V10A (108%), G11A (91%), G12A (31%), T13A (118%), N15A (68%), T16A (55%), P17A (83%), G18A (83%), T20A (131%), S22A (118%), V25A (133%), T27A (119%), R28A (100%), N29A (102%)
AssayType Insecticidal

Simonsen SM, Sando L, Rosengren KJ, Wang CK, Colgrave ML, Daly NL, Craik DJ (2008) Alanine scanning mutagenesis of the prototypic cyclotide reveals a cluster of residues essential for bioactivity J Biol Chem 283:9805-9813

Proteins Assayed [G6A]kalata B1
[E7A]kalata B1
[G1A]kalata B1
[L2A]kalata B1
[P3A]kalata B1
[V4A]kalata B1
[T8A]kalata B1
[V10A]kalata B1
[G11A]kalata B1
[T13A]kalata B1
[N15A]kalata B1
[T16A]kalata B1
[G18A]kalata B1
[T20A]kalata B1
[S22A]kalata B1
[V25A]kalata B1
[T27A]kalata B1
[R28A]kalata B1
[N29A]kalata B1
kalata B1
[G12A]kalata B1
[P17A]kalata B1