Evidence for membrane-bound oligomerization of bacteriophage phi X174 lysis protein-E.
The expression of cloned bacteriophage phi X174 lysis gene E was analyzed in minicells of Escherichia coli using two-dimensional gel electrophoresis. Beside the 10-11-kDa protein-E, at least two additional protein bands were detected, associated with the inner membrane, which showed the same isoelectric point as E. To clarify whether these proteins were E-specific, two different antibodies directed against a beta-galactosidase-E' hybrid protein and a synthetic oligopeptide corresponding to the C-terminal end of protein-E were raised. Immunoadsorption studies with anti-peptide-specific antibodies resulted in the detection of protein-E as well as in the detection of proteins of higher molecular weight. Two of these protein bands were positively recognized by anti beta-galactosidase-E' antibodies. The latter protein bands had the same molecular weight as the putative protein-E bands detected by two-dimensional gel electrophoresis indicating that these bands represent protein-E-specific oligomers. These data support the idea that an E-specific oligomeric structure penetrating the inner and outer membrane of E. coli is formed during the lytic action of protein-E.
Bläsi
U
U
Institute of Genetics and Microbiology, University of Munich, Federal Republic of Germany.
Linke
R P
RP
Lubitz
W
W
D000906:Antibodies / Q000276:immunology; D000941:Antigens / Q000276:immunology; D001426:Bacterial Proteins / Q000235:genetics / Q000276:immunology / Q000378:metabolism*; D010584:Bacteriophage phi X 174 / Q000378:metabolism*; D002462:Cell Membrane / Q000378:metabolism; D004279:DNA, Viral / Q000235:genetics; D015180:Electrophoresis, Gel, Two-Dimensional; D000939:Epitopes / Q000276:immunology; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism; D005786:Gene Expression Regulation; D007163:Immunosorbent Techniques; D046911:Macromolecular Substances; D008970:Molecular Weight; D009842:Oligopeptides / Q000276:immunology; D010957:Plasmids; D011401:Promoter Regions, Genetic; D011993:Recombinant Fusion Proteins / Q000276:immunology; D014169:Transformation, Bacterial; D014764:Viral Proteins / Q000235:genetics / Q000378:metabolism; D001616:beta-Galactosidase
D000906:Antibodies; D000941:Antigens; D001426:Bacterial Proteins; D004279:DNA, Viral; C029555:E protein, bacteriophage X174; D000939:Epitopes; D046911:Macromolecular Substances; D009842:Oligopeptides; D011993:Recombinant Fusion Proteins; D014764:Viral Proteins; D001616:beta-Galactosidase