Recognition of the colicin A N-terminal epitope 1C11 in vitro and in vivo in Escherichia coli by its cognate monoclonal antibody.
We demonstrate that the 1C10 monoclonal antibody (mAb) directed against the N-terminal domain of the colicin A recognizes a 13 residue-region (13Thr-Gly-Trp-Ser-Ser-Glu-Arg-Gly-Ser-Gly-Pro- Asp-Pro25). When this peptide is inserted into a protein in the amino-terminal or an internal position, the tagged protein is efficiently detected by the 1C11 mAb either by immunoblotting or immunoprecipitation. In vitro, the minimal structure required for detection using the pepscan system is 19Arg-Gly-Ser-Gly-Pro-Glu-Pro25, indicating that in vivo the proper exposure of the epitope requires additional residues. The construction of a versatile vector allowing overproduction of tagged proteins is described. Various applications of the 1C11 epitope are mentioned. This epitope did not alter the function of any of the proteins so far tested.
Geli
V
V
Laboratoire d'Ingénierie et Dynamique des Systèmes Membranaires, C.N.R.S., Marseille, France.
Lloubes
R
R
Zaat
S A
SA
van Spaendonk
R M
RM
Rollin
C
C
Benedetti
H
H
Lazdunski
C
C
D000595:Amino Acid Sequence; D000911:Antibodies, Monoclonal / Q000276:immunology; D001483:Base Sequence; D003001:Cloning, Molecular; D003087:Colicins / Q000235:genetics / Q000276:immunology*; D004269:DNA, Bacterial; D000939:Epitopes / Q000276:immunology*; D004926:Escherichia coli / Q000235:genetics / Q000276:immunology*; D005822:Genetic Vectors; D015151:Immunoblotting; D008969:Molecular Sequence Data
D000911:Antibodies, Monoclonal; D003087:Colicins; D004269:DNA, Bacterial; D000939:Epitopes