Part:BBa_K4165006
TD28rev (Tau binding peptide)
A synthetic peptide that is used for targeting misfolded tau protein (BBa_K4165009) as it binds to PHF6 of tau fibrils.
Usage and Biology
This is the reversed sequence of peptide TD28 (BBa_K4165145) which is a short synthetic peptide acquired by mirror image phage display for binding to tau fibrils at its seed sequence ‘VQIVYK’ called the PHF6. The aggregation of microtubule-associated tau protein starts at this seed to form neurofibrillary tangles inside the brain, which are the main cause of Alzheimer’s disease (AD). Accordingly, this peptide would be suitable to act as our targeting domain for the aggregations.
Sequence and features
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000COMPATIBLE WITH RFC[1000]
Dry Lab characterization
Modeling
We modeled the peptide with (Alphafold2, Apptest, iTASSER, and TRrosetta) and our top model was obtained from Apptest scoring 6 out of 6 according to our quality assessment code.
Figure 1.: Predicted 3D structure of TD28REV peptide visualized by Pymol.
Docking
Docking was performed using three different algorithms (Lightdock, Cluspro, and Galaxy web server) to test the interaction and binding affinity of TD28REV to both tau PHF filament and amyloid beta aggregates, this was followed by ranking of docked models and selection of models with the lowest free energy of binding, please find the code for docking and ranking at Programming club page.. .TD28REV vs Tau seed (PHF):
ΔG = -170.825
Figure 2.: Visualization of TD28REV docked with Tau PHF visualized by Pymol.
TD28REV vs Amyloid beta:
ΔG = -40.34
Figure 3.: Visualization of TD28REV docked with Amyloid beta visualized by Pymol.
Mathematical modeling
Transcription rate and translation rate under T7 promotor
the mathematical modeling was based on our code for the calculation of transcription and translation (you can find it in Programming club page.. besides the estimated results from the wet lab.
Figure 4. this figure shows the results from the transcription and translation code showing the
mRNA and protein concentrations variation with time compared with the wet lab results.
References
Dammers, C., Yolcu, D., Kukuk, L., Willbold, D., Pickhardt, M., Mandelkow, E., ... & Funke, S. A. (2016). Selection and Characterization of Tau Binding ᴅ-Enantiomeric Peptides with Potential for Therapy of Alzheimer Disease. PLoS One, 11(12), e0167432.
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