Translocation of inserted foreign epitopes by a channel-forming protein.
Certain bacterial protein toxins are able to insert themselves into, and at least partially across, lipid bilayer membranes in the absence of any auxiliary proteins, by using unknown mechanisms to overcome the high energy barrier presented by the hydrophobic bilayer core. We have previously shown that one such toxin, colicin Ia, translocates a large, hydrophilic part of itself completely across a lipid bilayer in conjunction with the formation of an ion-conducting channel. To address the question of whether the colicin can translocate any arbitrary amino acid sequence, we have altered the translocated segment by inserting, singly, two different foreign epitopes. Colicins containing either epitope retain significant bactericidal activity and form channels of normal conductance in planar bilayers. Furthermore, antibodies added on the side of the bilayer opposite that to which the colicin was added interact specifically with the corresponding epitopes, producing an inhibition of channel closing. Thus, the inserted epitopes are translocated along with the rest of the segment, suggesting that a surprisingly small part of colicin Ia, located elsewhere in the molecule, acts as a nonspecific protein translocator.
Jakes
K S
KS
Department of Physiology and Biophysics, Albert Einstein College of Medicine, 1300 Morris Park Avenue, Bronx, NY 10461, USA. jakes@aecom.yu.edu
Kienker
P K
PK
Slatin
S L
SL
Finkelstein
A
A
D000595:Amino Acid Sequence; D001483:Base Sequence; D003087:Colicins / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism*; D000939:Epitopes; D004926:Escherichia coli / Q000378:metabolism; D006388:Hemagglutinins; D007473:Ion Channels / Q000737:chemistry*; D008051:Lipid Bilayers; D008969:Molecular Sequence Data; D016254:Mutagenesis, Insertional / Q000379:methods*; D009838:Oligodeoxyribonucleotides; D009842:Oligopeptides; D010455:Peptides; D011994:Recombinant Proteins / Q000737:chemistry / Q000378:metabolism
D003087:Colicins; D000939:Epitopes; D006388:Hemagglutinins; D007473:Ion Channels; D008051:Lipid Bilayers; D009838:Oligodeoxyribonucleotides; D009842:Oligopeptides; D010455:Peptides; D011994:Recombinant Proteins; C085706:FLAG peptide