BIOL 156 |
| The Clostridium neurotoxin family includes 7 serotypes of botulinum neurotoxin (BoNT/A-/G), and a single tetanus neurotoxin (TeNT). BoNT/A, /C and /E cleave SNAP-25. BoNT/B, /D, /F, /G and TeNT cleave the VAMP. To understand how the specificity of molecular recognition between the neurotoxin and their substrates evolved and the positional significance of protein sequence of substrates vs. the neurotoxin enzyme activity, we decided to conjugate a green fluorescence protein to the C-terminus of SNAP-25 to improve the resolution of both densitometry-based cleavage activity assay (SDS-PAGE) and immunofluorescence-based functional assay (ELISA). Our result has shown the EGFP labeled SNAP-25, can be cleaved by recombinant BoNT/A and BoNT/E light chains effectively. The chimerical construction of the substrate does not affect the neurotoxin activity. This result encourages us to conjugate another fluorescence protein to the N-terminus of substrate to construct the Fluorescence-Resonance-Energy-Transfer system to be used in the neurotoxin detection in the future. Funded by DoD/Army Contract (W911NF-06-1-0095) |
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Frontiers in Chemical Biology
5:00 PM-7:00 PM, Wednesday, August 22, 2007 BCEC -- Exhibit Hall - B2, Poster
Division of Biological Chemistry |