title: Fine Mutational Analysis of 2B8 and 3H7 Tag Epitopes with Corresponding Specific Monoclonal Antibodies.
abstract: Bacteriophytochromes are phytochrome-like light-sensing photoreceptors that use biliverdin as a chromophore. To study the biochemical properties of the Deinococcus radiodurans bacteriophytochrome (DrBphP) protein, two anti-DrBphP mouse monoclonal antibodies (2B8 and 3H7) were generated. Their specific epitopes were identified in our previous report. We present here fine epitope mapping of these two antibodies by using truncation and substitution of original epitope sequences in order to identify minimized epitope peptides. The previously reported original epitope sequences for 2B8 and 3H7 were truncated from both sides. Our analysis showed that the minimal peptide sequence lengths for 2B8 and 3H7 antibodies were nine amino acids (RDPLPFFPP) and six amino acids (PGEIEE), respectively. We further characterized these peptides in order to investigate their reactivity after single deletion and single substitution of the original peptides. We found that single-substituted 2B8 epitope (RDPLPAFPP) and dual-substituted 3H7 epitope (PGEIAD) showed significantly increased reactivity. These two antibodies with high reactivity for the short modified peptide sequences are valueble for developing new peptide tags for protein research.
authors.affiliation: Graduate School of Biotechnology, Kyung Hee University, Yongin 17104, Korea.
authors.forename: Tae-Lim
authors.initials: TL
authors.lastname: Kim
authors.affiliation: Graduate School of Biotechnology, Kyung Hee University, Yongin 17104, Korea.
authors.forename: Man-Ho
authors.initials: MH
authors.lastname: Cho
authors.affiliation: Graduate School of Biotechnology, Kyung Hee University, Yongin 17104, Korea.
authors.forename: Kanidta
authors.initials: K
authors.lastname: Sangsawang
authors.affiliation: Graduate School of Biotechnology, Kyung Hee University, Yongin 17104, Korea.
authors.forename: Seong Hee
authors.initials: SH
authors.lastname: Bhoo
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D000911:Antibodies, Monoclonal / Q000378:metabolism*; D000918:Antibody Specificity; D001426:Bacterial Proteins / Q000737:chemistry / Q000235:genetics / Q000276:immunology*; D004252:DNA Mutational Analysis; D034301:Deinococcus / Q000235:genetics / Q000276:immunology / Q000378:metabolism*; D018604:Epitope Mapping; D000939:Epitopes / Q000235:genetics* / Q000276:immunology; D051379:Mice
chemical_list: D000911:Antibodies, Monoclonal; D001426:Bacterial Proteins; D000939:Epitopes
keywords: 2B8 monoclonal antibody; 3H7 monoclonal antibody; DrB-phP protein; epitope mapping; tag