A pH-induced increase in hydrophobicity as a possible step in the penetration of colicin E3 through bacterial membranes.
Exposure to low pH triggers an increase in the hydrophobicity of the colicin E3 molecule. Using a [3H] Triton X-100 binding assay we have shown that the amount of detergent (at supramicellar concentrations) associated with colicin E3 increased dramatically at pH 3.8 and below. Interaction of colicin E3 with asolectin vesicles was monitored by following its cross-linking with two different photoactivatable radioactive phospholipid analogues. At neutral pH colicin E3 was cross-linked with the phospholipid probing the membrane surface whereas at pH 4.5 and below, the bacteriocin reacted with the phospholipid probing the hydrophobic core of the bilayer. With the use of phase partitioning of proteins in Triton X-114 it was shown that at acidic pH whole colicin E3 and its immunity protein segregated in the detergent phase. After trypsin digestion of the colicin-immunity complex, the N-terminal portion of E3 (T1) and the immunity partitioned in the detergent phase at low pH. In contrast, the enzymic domain of the colicin (T2) remained in the aqueous phase and was recovered in a highly active form as a consequence of its dissociation from the immunity protein. These results are discussed in relation to the mechanism of entry of colicin E3 into bacterial cells.
D002250:Carbon Radioisotopes; D002462:Cell Membrane / Q000378:metabolism*; D003087:Colicins / Q000302:isolation & purification / Q000378:metabolism*; D003902:Detergents / Q000378:metabolism; D004926:Escherichia coli / Q000378:metabolism*; D006863:Hydrogen-Ion Concentration; D007700:Kinetics; D008051:Lipid Bilayers; D017830:Octoxynol; D010713:Phosphatidylcholines; D010743:Phospholipids / Q000378:metabolism; D011092:Polyethylene Glycols / Q000378:metabolism; D014316:Tritium; D014357:Trypsin
D002250:Carbon Radioisotopes; D003087:Colicins; D003902:Detergents; D008051:Lipid Bilayers; D010713:Phosphatidylcholines; D010743:Phospholipids; D014316:Tritium; D011092:Polyethylene Glycols; C024990:asolectin; D017830:Octoxynol; D014357:Trypsin