Preparation and characterization of two monoclonal antibodies against different epitopes in Escherichia coli ribosomal protein L7/L12.
Two monoclonal antibodies with specificities for Escherichia coli 50 S ribosomal subunit protein L7/L12 were isolated. The antibodies and Fab fragments thereof were purified by affinity chromatography using solid-phase coupled L7/L12 protein as the immunoadsorbent. The two antibodies were shown to recognize different epitopes; one in the N-terminal and the other in the C-terminal domain of protein L7/L12. Both intact antibodies strongly inhibited polyuridylic acid-directed polyphenylalanine synthesis, ribosome-dependent GTPase activity, and the binding of elongation factor EF-G to the ribosome. Ratios of antibody to ribosome of 4:1 or less were effective in inhibiting these activities. Neither antibody prevented the association of ribosomal subunits to form 70 S ribosomes. The Fab fragments showed similar effects.
Sommer
A
A
Etchison
J R
JR
Gavino
G
G
Zecherle
N
N
Casiano
C
C
Traut
R R
RR
D000818:Animals; D000911:Antibodies, Monoclonal; D002846:Chromatography, Affinity; D004591:Electrophoresis, Polyacrylamide Gel; D000939:Epitopes / Q000276:immunology*; D004926:Escherichia coli; D020558:GTP Phosphohydrolases / Q000378:metabolism; D051379:Mice; D008807:Mice, Inbred BALB C; D010452:Peptide Biosynthesis; D010455:Peptides; D011072:Poly U / Q000378:metabolism; D012269:Ribosomal Proteins / Q000276:immunology*
D000911:Antibodies, Monoclonal; D000939:Epitopes; D010455:Peptides; D012269:Ribosomal Proteins; D011072:Poly U; C012001:polyphenylalanine; C016700:ribosomal protein L7-L12; D020558:GTP Phosphohydrolases