Difference between revisions of "Part:BBa K4165009"

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Tau (0N4R)

This basic part encodes the human microtubule-associated tau protein isoform 0N4R.

Usage and Biology

Alzheimer's disease (AD), which is considered the most common neurodegenerative disease to cause dementia, is characterized by 2 main accumulations that are amyloid plaques from amyloid beta and NFTs aggregates resulting from hyperphosphorylated or abnormally phosphorylated tau protein accumulation. Our part Tau, microtubule-associated protein (MAP) is a phosphoprotein that is prevalently found in cytosol and neuron axons. It is determined to be significantly expressed in neurons of the central nervous system (CNS) and the ocular tissues. It has a crucial role both under normal physiological conditions and also in the pathology of Alzheimer's disease.

In the normal brain, it can stabilize the neuronal microtubules that are essential for the establishment of cell polarity, the development of cell processes, and intracellular signal transduction. Six molecular tau isoforms are coded by a single gene on chromosome 17 resulting from alternative splicing of tau pre-mRNA and characterized to be significantly hydrophilic, heat stable, and soluble. These six isoforms differ in their binding repeats either 3R taus or 4R taus microtubule-binding repeats and the extra 4R repeat comes from the second (R2) repeat found in 4R. Tau biological activity is affected by 2 main processes that are alternative splicing and phosphorylation. For tau's interaction with tubulin and the enhancement of microtubule assembly, normal brain tau appears to require 2-3 moles of phosphate per mole of the protein. However, tau is phosphorylated at Ser262 and Ser214 in AD, which causes tau to separate from microtubules.

In the AD brain, tau Hyperphosphorylation is considered the main cause of AD progression, it may alter the protein's shape and charge, which in turn causes the microtubule-binding domain to become exposed and allow tau to self-assemble and form oligomers characterized to be neurofibrillary tangle. According to several studies, the polymerized tau (neurofibrillary tangles) is inert since it does not bind to tubulin or encourage its assembly into microtubules.

Before the development of NFTs, all six forms of tau are self-assembled into paired helical filaments as a result of hyperphosphorylation at the C-terminus of tau (PHFs). The aggregated tau protein takes this shape, which impairs axonal transit and continuously promotes microtubule instability. AD patients have an aberrant or hyperphosphorylated tau protein concentration that is four times higher than that of normal controls. These misfolded tau proteins are also recognized as potential neurotoxins and lose their basic function of MT stability along with enhanced aggregation effects.

Sequence and Features

Dry Lab Characterization

Modeling

The structure of Tau was modeled by several tools and the top model was retrieved from trRosetta ranking 5 out of 6 according to our quality assessment code.

                              Figure 1. Predicted 3D structure of Tau(0N4R) modeled by trRosetta

WetLab Results

Transformation of His Tau in BL-21 using pGS-21a vector

                                    Figure 2. Transformed plate of His Tau + pGS-21a

Transformation of His Tau in DH-5 alpha using pJET vector

                                   Figure 3. Transformed plate of His Tau + pJET

SDS PAGE of induced and non induced samples of His Tau

            Figure 4. This figure shows the comparison between the induced and non-induced samples of His Tau, 
            where well no.2 is the non-induced sample while well no.5 is the induced sample showing that our 
            protein is induced effectively owing to our right choice of IPTG, time interval and concentration

References

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2- Muralidar, S., Ambi, S., Sekaran, S., Thirumalai, D. and Palaniappan, B., 2020. Role of tau protein in Alzheimer's disease: The prime pathological player. International Journal of Biological Macromolecules, 163, pp.1599-1617.

3- R. Sajjad, R. Arif, A.A. Shah, I. Manzoor, G. Mustafa Pathogenesis of Alzheimer’s disease: role of amyloid-β and hyperphosphorylated tau protein Indian J. Pharm. Sci., 80 (2018), pp. 581-591, 10.4172/pharmaceutical-sciences.1000397

4- Köpke, E., Tung, Y., Shaikh, S., Alonso, A., Iqbal, K. and Grundke-Iqbal, I., 1993. Microtubule-associated protein tau. Abnormal phosphorylation of a non-paired helical filament pool in Alzheimer disease. Journal of Biological Chemistry, 268(32), pp.24374-24384.

5- Iqbal, K., Gong, C. and Liu, F., 2013. Hyperphosphorylation-Induced Tau Oligomers. Frontiers in Neurology, 4.

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8- Pérez, M., Cuadros, R., Smith, M., Perry, G. and Avila, J., 2000. Phosphorylated, but not native, tau protein assembles following reaction with the lipid peroxidation product, 4-hydroxy-2-nonenal. FEBS Letters, 486(3), pp.270-274.

9- Moszczynski, A., Gohar, M., Volkening, K., Leystra-Lantz, C., Strong, W. and Strong, M., 2015. Thr175-phosphorylated tau induces pathologic fibril formation via GSK3β-mediated phosphorylation of Thr231 in vitro. Neurobiology of Aging, 36(3), pp.1590-1599.

10- Lee, H., Perry, G., Moreira, P., Garrett, M., Liu, Q., Zhu, X., Takeda, A., Nunomura, A. and Smith, M., 2005. Tau phosphorylation in Alzheimer's disease: pathogen or protector?. Trends in Molecular Medicine, 11(4), pp.164-169.

11- Chu, D. and Liu, F., 2018. Pathological Changes of Tau Related to Alzheimer’s Disease. ACS Chemical Neuroscience, 10(2), pp.931-944.

12- Lin, Y., Cheng, J., Liang, L., Ko, C., Lo, Y. and Lu, P., 2007. The binding and phosphorylation of Thr231 is critical for Tau’s hyperphosphorylation and functional regulation by glycogen synthase kinase 3β. Journal of Neurochemistry, 103(2), pp.802-813.

13J. Neddens, M. Temmel, S. Flunkert, B. Kerschbaumer, C. Hoeller, T. Loeffler, V. Niederkofler, G. Daum, J. Attems, B. Hutter-Paier

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18- Braak, H., Braak, E., Grundke-Iqbal, I., & Iqbal, K. (1986). Occurrence of neuropil threads in the senile human brain and in Alzheimer's disease: A third location of paired helical filaments outside of neurofibrillary tangles and neuritic plaques. Neuroscience Letters, 65(3), 351-355. doi: 10.1016/0304-3940(86)90288-0

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21- Miao, J., Shi, R., Li, L., Chen, F., Zhou, Y., & Tung, Y. et al. (2019). Pathological Tau From Alzheimer’s Brain Induces Site-Specific Hyperphosphorylation and SDS- and Reducing Agent-Resistant Aggregation of Tau in vivo. Frontiers In Aging Neuroscience, 11. doi: 10.3389/fnagi.2019.00034

22- DeTure, M., & Dickson, D. (2019). The neuropathological diagnosis of Alzheimer’s disease. Molecular Neurodegeneration, 14(1). doi: 10.1186/s13024-019-0333-5

23- Uddin, M., Ashraf, G., Mamun, A., & Mathew, B. (2020). Toxic tau: structural origins of tau aggregation in Alzheimer's disease. Neural Regeneration Research, 15(8), 1417. doi: 10.4103/1673-5374.274329