%0 Figure %A J. Dowd, Kelley %A Tweten, Rodney K. %D 2013 %T TMH insertion in PFO and PFOR468A. %U https://plos.figshare.com/articles/figure/_TMH_insertion_in_PFO_and_PFO_R468A_/284497 %R 10.1371/journal.ppat.1002787.g006 %2 https://ndownloader.figshare.com/files/613997 %K insertion %K pfo %X

A cysteine was substituted for Ala-215 in PFO and PFOR468A, which is located in TMH1.The sidechain of Ala-215 is in an aqueous environment in the soluble monomer, but enters the membrane upon formation of the membrane spanning β-barrel [27]. Therefore an NBD probe positioned at this site undergoes a polar to nonpolar transition that is detected by an increase in the fluorescence emission of the probe.(A) Each NBD-labeled derivative was incubated in the presence (dashed line) and absence (solid line) of human erythrocyte ghost membranes. (B) Unlabeled native PFO or PFOR468A were mixed in a 4∶1 molar ratio with PFOA215C-NBD and PFOR468A•A215C-NBD derivatives. The fluorescence emission for all experiments wasrelative to the maximum emission change observed for NBD-labeled PFO. The fluorescence emission intensity of NBD was measured from 500–600 nm. The data are representative of 3 experiments.

%I PLOS Pathogens