Individual domains of colicins confer specificity in colicin uptake, in pore-properties and in immunity requirement.
Six different hybrid colicins were constructed by recombining various domains of the two pore-forming colicins A and E1. These hybrid colicins were purified and their properties were studied. All of them were active against sensitive cells, although to varying degrees. From the results, one can conclude that: (1) the binding site of OmpF is located in the N-terminal domain of colicin A; (2) the OmpF, TolB and TolR dependence for translocation is also located in this domain; (3) the TolC dependence for colicin E1 is located in the N-terminal domain of colicin E1; (4) the 183 N-terminal amino acid residues of colicin E1 are sufficient to promote E1AA uptake and thus probably colicin E1 uptake; (5) there is an interaction between the central domain and C-terminal domain of colicin A; (6) the individual functioning of different domains in various hybrids suggests that domain interactions can be reconstituted in hybrids that are fully active, whereas in others that are much less active, non-proper domain interactions may interfere with translocation; (7) there is a specific recognition of the C-terminal domains of colicin A and colicin E1 by their respective immunity proteins.
Benedetti
H
H
Centre de Biochimie et de Biologie Moléculaire de C.N.R.S., Marseille, France.
Frenette
M
M
Baty
D
D
Knibiehler
M
M
Pattus
F
F
Lazdunski
C
C
D001425:Bacterial Outer Membrane Proteins; D001692:Biological Transport; D003087:Colicins / Q000737:chemistry / Q000378:metabolism*; D004553:Electric Conductivity; D004926:Escherichia coli / Q000502:physiology; D029968:Escherichia coli Proteins; D006863:Hydrogen-Ion Concentration; D007473:Ion Channels / Q000502:physiology*; D008051:Lipid Bilayers; D008564:Membrane Potentials; D026901:Membrane Transport Proteins; D011956:Receptors, Cell Surface / Q000502:physiology; D018000:Receptors, Peptide; D011993:Recombinant Fusion Proteins / Q000378:metabolism; D013329:Structure-Activity Relationship; D014882:Water-Electrolyte Balance
D001425:Bacterial Outer Membrane Proteins; C044592:BtuB protein, E coli; D003087:Colicins; D029968:Escherichia coli Proteins; D007473:Ion Channels; D008051:Lipid Bilayers; D026901:Membrane Transport Proteins; D011956:Receptors, Cell Surface; D018000:Receptors, Peptide; D011993:Recombinant Fusion Proteins