Characterization of Variants of the Pore-Forming Toxin ClyA from Escherichia coli Controlled by a Redox Switch.

Cytotoxin ClyA,Hemolysis-inducing protein,Latent pore-forming 34 kDa hemolysin,Silent hemolysin SheA

TEIVCDKTVEVVKNAIETADGALDLYNKYLDQVIPWQTFDETIKELSRFKQEYSQAASVLVGDIKTLLMDSQDKYFEATQ
TVYEWAGVATQLLAAYILLFDEYNEKKASAQKDILIKVLDDGITKLNEAQKSLLVSSQSFNNASGKLLALDSQLTNDFSE
KSSYFQSQVDKIRKEAYAGAAAGVVAGPFGLIISYSIAAGVVEGKLIPELKNKLKSVQNFFTTLSNTVKQANKDIDAAKL
KLTTEIAAIGEIKTETETTRFYCDYDDLMLSLLKEAAKKMINTANEYQKRHGKKTLFEVPEV

4 Quick-change PCRs to introduce point mutations, TEV cleavage of N-terminal His(6)-tag
hlyE, clyA, hpr, sheA, ycgD, b1182, JW5181

alpha pore-forming toxin, intramolecular disulfide bond, toxin

TEIVADKTVEVVKNAIETADGALDLYNKYLDQVIPWQTFDETIKELSRFKQEYSQAASVLVGDIKTLLMDSQDKYFEATQ
TVYEWCGVATQLLAAYILLFDEYNEKKASAQKDILIKVLDDGITKLNEAQKSLLVSSQSFNNASGKLLALDSQLTNDFSE
KSSYFQSQVDKIRKEAYAGAAAGVVAGPFGLIISYSIAAGVVEGKLIPELKNKLKSVQNFFTTLSNTVKQANKDIDAAKL
KLTTEIAAIGEIKTETETTRFYVDYDDLMLSLLKEAAKKMINTCNEYQKRHGKKTLFEVPEV