• Journal of Applied Biological Chemistry
• /
• v.53 no.2
• /
• pp.112-115
• /
• 2010

To analyze the surface properties of bacteriophytochrome (BphP), five (2B8, 2C11, 3B2, 3D2, 3H7) anti-BphP monoclonal antibodies were produced by using full-length of BphP of Deinococcus radiodurnas. 2B8 and 2C11 preferentially recognized the epitopes at N-terminal region of BphP, whereas 3B2, 3D2 and 3H7 showed preferential affinities to the epitopes of C-terminal region of BphP.

• Molecules and Cells
• /
• v.39 no.6
• /
• pp.460-467
• /
• 2016

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.