Structural and functional properties of colicin B.
Colicin B was isolated in pure form from cells of Escherichia coli that contained the colicin activity and immunity genes cloned on a multi-copy plasmid. Active colicin B consisted of a single polypeptide with Mr of about 60,000. The sequence of 44 amino acids from the amino-terminal portion is presented. The isoelectric point of the protein was at 4.5. Colicin B inhibited the membrane potential-dependent transport of proline and enhanced the uptake of alpha-methylglucoside via the phosphoenolpyruvate-dependent phosphotransferase system. Colicin B formed small, ion permeable channels with an average single-channel conductance of 13.7 pS (1 pS = 10(-12) siemens) in 1 M KCl. Channel formation was voltage-dependent in the pH range between 4.5 and 6. At pH 7 the channels were voltage independent. Voltage-dependent channels were only formed when the trans compartment (the protein was added to the cis compartment) was negative by at least 70 mV. Evidence for an asymmetric single channel conductance was obtained. With KCl a hyperbolic conductance-concentration relationship was observed. The conductance for monovalent cations was minimal for Li+ and was maximal for NH+4. The single channel conductance of colicin B was larger than that of colicin A as judged from lipid bilayer experiments under otherwise identical conditions.
U
U
Pressler
V
V
Braun
B
B
Wittmann-Liebold
R
R
Benz
D000595:Amino Acid Sequence; D026921:Amino Acid Transport Systems, Neutral; D000641:Ammonia / Q000494:pharmacology; D002352:Carrier Proteins / Q000037:antagonists & inhibitors; D003087:Colicins / Q000378:metabolism*; D004553:Electric Conductivity; D004591:Electrophoresis, Polyacrylamide Gel; D004926:Escherichia coli / Q000032:analysis; D029968:Escherichia coli Proteins; D007473:Ion Channels / Q000378:metabolism; D008433:Mathematics; D008757:Methylglucosides / Q000378:metabolism; D008970:Molecular Weight; D011189:Potassium Chloride / Q000494:pharmacology; D027981:Symporters
D026921:Amino Acid Transport Systems, Neutral; D002352:Carrier Proteins; D003087:Colicins; D029968:Escherichia coli Proteins; D007473:Ion Channels; D008757:Methylglucosides; D027981:Symporters; C041832:proline transport protein; C060609:PutP protein, E coli; C027020:methylglucoside; D011189:Potassium Chloride; D000641:Ammonia