[E7A]kalata B1(459) Protein Card

General Information
Name [E7A]kalata B1
Sequence GLPVCGATCVGGTCNTPGCTCSWPVCTRN
Class Cyclotide
Average Mass 2834.24
Monoisotopic Mass 2832.14
m/z M+H 2833.14
ProteinType Mutant
Parent kalata B1
Organism Synthetic
Notes
Cyclic Yes

Assay
Insecticidal
Substitution of certain residues of kalata B1 to alanine result in loss of insecticidal activity [...]Simonsen SM et al. (2008) J Biol Chem 283:9805-9813
Hemolytic
Substitution of certain residues of kalata B1 to alanine result in loss of hemolytic activity. Residues in loops 1 and 3 along with a few other isolated residues strongly affect activity. [...]Simonsen SM et al. (2008) J Biol Chem 283:9805-9813
Metal binding assay
In the presence of Mn2+ kalata B1, T16A and T27A showed specific broadening of a defined set of amide signals. There was almost complete broadening of the amide signals from C14, N15, and T16/A16, and significant broadening (i.e. ~70% attenuation) of loop 1 residues. By contrast, E7A showed significantly less broadening. [...]Simonsen SM et al. (2008) J Biol Chem 283:9805-9813
Temperature
G6A and E7A stable in high temperature. [...]Simonsen SM et al. (2008) J Biol Chem 283:9805-9813
Acid Hydrolysis
G6A and E7A showed no degradation at 37 C for up to 24 h. At 80 C, they degraded within 90 min. [...]Simonsen SM et al. (2008) J Biol Chem 283:9805-9813
Enzymatic Digest
G6A and E7A mutants of kalata B1 were completely impervious to enzymatic digestion by the three endoproteinases trypsin, chymotrypsin, and proteinase K, as was the prototypic cyclotide kalata B1. [...]Simonsen SM et al. (2008) J Biol Chem 283:9805-9813
Nematocidal
The cyclotides caused a concentration-dependent response in the LDA with the two nematode species. [...]Colgrave ML et al. (2008) Biochemistry 47:5581-5589

References
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

Cross-references
Nucleic acids
Structure Solution structure of [E7A]kalata B1
Links

Tools