Sequence analysis, expression, and paratope characterization of a single-chain Fv fragment for the eukaryote ribosomal P proteins.
The variable genes of monoclonal antibody (mAb) B10, specific for the C-terminal region of the eukaryotic ribosomal P protein, have been cloned as a single-chain Fv fragment (scFv) and expressed in Escherichia coli. The primary sequence of the variable regions of the B10 antibody, together with a detailed characterization of the reactive residues of the antigen, allowed the construction of a model of the paratope-epitope interaction, giving a first insight into the binding mechanisms of anti-P autoantibodies to their target peptides. The mAb and scFv could be useful for extensive P protein detection since both recognize the highly conserved motif DDxGF.
López Bergami
Pablo
P
Instituto de Biología y Medicina Experimental, Vuelta de Obligado 2490 (1428), Buenos Aires, Argentina.
Mateos
Pablo
P
Hoebeke
Johan
J
Levin
Mariano Jorge
MJ
Baldi
Alberto
A
D020816:Amino Acid Motifs; D000595:Amino Acid Sequence; D000818:Animals; D000911:Antibodies, Monoclonal / Q000737:chemistry / Q000235:genetics; D001666:Binding Sites, Antibody / Q000235:genetics; D004247:DNA / Q000235:genetics; D005803:Genes, Immunoglobulin; D007135:Immunoglobulin Variable Region / Q000737:chemistry / Q000235:genetics*; D051379:Mice; D008958:Models, Molecular; D008969:Molecular Sequence Data; D015800:Protozoan Proteins / Q000276:immunology; D012269:Ribosomal Proteins / Q000276:immunology*; D017386:Sequence Homology, Amino Acid; D014349:Trypanosoma cruzi / Q000276:immunology
D000911:Antibodies, Monoclonal; D007135:Immunoglobulin Variable Region; C064601:L12E protein, Trypanosoma cruzi; D015800:Protozoan Proteins; D012269:Ribosomal Proteins; D004247:DNA