Conformational changes of urea-denatured colicin E1 induced by phospholipid membranes.
The membrane insertion of urea-denatured colicin E1 was studied by using fluorescence spectroscopy, circular dichroism and monolayer techniques. The results showed that the denatured colicin E1 taking mainly the 'random coil' conformation may recover its orderliness to a certain extent under the induction of the phospholipid membrane and insert spontaneously into phospholipid membrane, indicating that unfolding of colicin E1 does not inhibit its membrane insertion. Among the four tryptophan residues of the membrane-bound colicin E1 molecules, at least two were accessible by the quenchers, i.e. not inserted into the membranes. Although urea-denatured colicin E1 interacted preferentially with negatively charged phospholipids, it seems less dependent on the negatively charged lipid than colicin A. The addition of urea increased the speed of the adsorption of colicin E1 to the membrane, but did not affect obviously its membrane insertion ability.
Wu
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Department of Biological Sciences and Biotechnology, Tsinghua University, Beijing, People's Republic of China.
Sui
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SF
D055598:Chemical Phenomena; D002627:Chemistry, Physical; D002942:Circular Dichroism; D003087:Colicins / Q000737:chemistry / Q000187:drug effects*; D004134:Dimyristoylphosphatidylcholine / Q000494:pharmacology; D008081:Liposomes; D008563:Membrane Lipids / Q000494:pharmacology*; D010714:Phosphatidylethanolamines / Q000494:pharmacology; D010715:Phosphatidylglycerols / Q000494:pharmacology; D010743:Phospholipids / Q000494:pharmacology*; D011487:Protein Conformation / Q000187:drug effects*; D011489:Protein Denaturation; D017510:Protein Folding; D013050:Spectrometry, Fluorescence; D013696:Temperature; D014364:Tryptophan / Q000737:chemistry; D014508:Urea / Q000494:pharmacology*
D003087:Colicins; D008081:Liposomes; D008563:Membrane Lipids; D010714:Phosphatidylethanolamines; D010715:Phosphatidylglycerols; D010743:Phospholipids; D014364:Tryptophan; D014508:Urea; C002773:dimyristoylphosphatidylglycerol; D004134:Dimyristoylphosphatidylcholine; C018525:1,2-dimyristoylphosphatidylethanolamine