Epitope mapping of Escherichia coli cell division protein FtsZ with monoclonal antibodies.
A fusion between lacZ and ftsZ of Escherichia coli was constructed to obtain a beta-galactosidase-FtsZ fusion protein. This fusion protein was used to raise antibodies against cell division protein FtsZ. Six monoclonal antibodies were obtained, and they reacted with FtsZ from cytoplasm and membrane fractions. The epitopes in FtsZ were localized by studying the reactions of the monoclonal antibodies with fusion proteins truncated at the carboxy terminus and with fragments that were obtained by CNBr cleavage of purified FtsZ. Five different epitopes were defined. Epitopes I and III reacted with the same monoclonal antibody, without showing apparent amino acid homology. Epitope II was defined by monoclonal antibodies that cross-reacted with an unknown cytoplasmic 50-kDa protein not related to FtsZ. Epitopes IV and V were recognized by different monoclonal antibodies. All monoclonal antibodies reacted strongly under native conditions, so it is likely that the five epitopes are situated on the surface of native FtsZ. By using these data and computer analysis, a provisional model of FtsZ is proposed. The FtsZ protein is considered to be globular, with a hydrophobic pocket containing GTP-binding elements. Epitopes I and II are situated on each side of the hydrophobic pocket. Because the carboxy terminus contains epitope V, the carboxy terminus of FtsZ is likely oriented toward the protein's surface.
Voskuil
J L
JL
Section of Molecular Cytology, University of Amsterdam, The Netherlands.
Westerbeek
C A
CA
Wu
C
C
Kolk
A H
AH
Nanninga
N
N
D000818:Animals; D000907:Antibodies, Bacterial / Q000276:immunology; D000911:Antibodies, Monoclonal / Q000276:immunology; D001426:Bacterial Proteins / Q000096:biosynthesis / Q000235:genetics / Q000276:immunology*; D002455:Cell Division; D003429:Cross Reactions; D003598:Cytoskeletal Proteins; D000939:Epitopes / Q000276:immunology*; D004926:Escherichia coli / Q000254:growth & development / Q000276:immunology*; D019204:GTP-Binding Proteins / Q000276:immunology*; D007074:Immunoglobulin G / Q000276:immunology; D007613:Kanamycin Resistance / Q000235:genetics; D051379:Mice; D008807:Mice, Inbred BALB C; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis / Q000276:immunology; D001616:beta-Galactosidase / Q000096:biosynthesis / Q000235:genetics
D000907:Antibodies, Bacterial; D000911:Antibodies, Monoclonal; D001426:Bacterial Proteins; D003598:Cytoskeletal Proteins; D000939:Epitopes; C071525:FtsZ protein, Bacteria; D007074:Immunoglobulin G; D011993:Recombinant Fusion Proteins; D001616:beta-Galactosidase; D019204:GTP-Binding Proteins