Interactions of colicin A domains with phospholipid monolayers and liposomes: relevance to the mechanism of action.
The colicin A polypeptide chain (592 amino acid residues) contains three domains which are linearly organized and participate in the sequential steps involved in colicin action. We have compared the penetrating ability in phospholipid monolayers and the ability to promote vesicle fusion at acidic pH of colicin A and of protein derivatives containing various combinations of its domains. The NH2-terminal domain (171 amino acid residues), required for translocation across the outer membrane, has little affinity for dilauroylphosphatidylglycerol (DLPG) monolayers at all pHs tested. The central domain has a pH-dependent affinity, although lower than that of the entire colicin A. The COOH-terminal domain contains a high-affinity lipid binding site, but in addition an electrostatic interaction is required as a first step in the process of penetration into negatively charged DLPG films. In contrast to the constructs containing the ionophoric domain, the NH2-terminal domain alone has no fusogenic activity for liposomes. These results are discussed with regard to the mechanism of entry and action of colicin A in sensitive cells. Our results suggest the existence of a pH-dependent interaction between the receptor binding domain (amino acid residues 172-388) and the pore-forming domain of colicin A (amino acid residues 389-592).
Frenette
M
M
Centre de Biochimie et de Biologie Moléculaire du CNRS, Marseille, France.
Knibiehler
M
M
Baty
D
D
Géli
V
V
Pattus
F
F
Verger
R
R
Lazdunski
C
C
D001665:Binding Sites; D003087:Colicins / Q000633:toxicity; D004735:Energy Transfer; D006863:Hydrogen-Ion Concentration; D007700:Kinetics; D008081:Liposomes; D008962:Models, Theoretical; D010713:Phosphatidylcholines; D010714:Phosphatidylethanolamines; D010715:Phosphatidylglycerols; D011312:Pressure; D011485:Protein Binding; D011487:Protein Conformation; D013329:Structure-Activity Relationship; D013499:Surface Properties
D003087:Colicins; D008081:Liposomes; D010713:Phosphatidylcholines; D010714:Phosphatidylethanolamines; D010715:Phosphatidylglycerols; C054349:1,2-didodecanoyl-glycero-3-phosphocholine