Abstract
More than half of the mutations associated with familiar Alzheimer’s disease have been found in the transmembrane domain of amyloid precursor protein (APP). These pathogenic mutations presumably influence the APP transmembrane domain structural and dynamic properties and result in its conformational change or/and lateral dimerization. Despite much data about the pathogenesis of Alzheimer’s disease, the initial steps of the pathogenesis remain unclear so far. For the investigation of the molecular basis of Alzheimer’s disease, we selected amyloid precursor protein fragment APP671-726 containing the transmembrane and metal-binding domains. This fragment is the substrate of the γ-secretase complex whose abnormal activity leads to the formation of amyloidogenic Aβ42 peptides. This work for the first time describes a highly effective cell-free APP671-726 production method and improved method of bacterial synthesis. Both methods yield milligram quantities of isotope-labeled protein for structural study by high resolution NMR spectroscopy in membrane mimicking milieus.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
Abbreviations
- AD:
-
Alzheimer’s disease
- APP:
-
amyloid precursor protein
- APPmc:
-
APP sequence limited by 671–726 residues (mc, metal chelating)
- APPtm:
-
APP sequence limited by 686–726 residues (tm, transmembrane)
- TM:
-
transmembrane
References
Grigorenko, A. P., and Rogaev, E. I. (2007) Mol. Biol. (Moscow), 41, 331–345.
Steiner, H., Fluhrer, R., and Haass, C. (2008) J. Biol. Chem., 283, 29627–29631.
Lovell, M. A., Robertson, J. D., Teesdale, W. J., Campbell, J. L., and Markesbery, W. R. (1998) J. Neurol. Sci., 158, 47–52.
Chen, W., Gamache, E., Richardson, D., Du, Zh., and Wang, C. (2012) Prot. Expr. Pur., 81, 11–17.
Gorman, P. M., Kim, S., Guo, M., Melnyk, R. A., McLaurin, J., Fraser, P. E., Bowie, J. U., and Chakrabartty, A. (2008) BMC Neurosci., 30, 9–17.
Sato, T., Tang, T. C., Reubins, G., Fei, J. Z., Fujimoto, T., Kienlen-Campard, P., Constantinescu, S. N., Octave, J. N., Aimoto, S., and Smith, S. O. (2009) PNAS, 106, 1421–1426.
Zhuang, T., Jap, B. K., and Sanders, C. R. (2011) J. Am. Chem. Soc., 133, 20571–20580.
Beel, A. J., Sakakura, M., Barrett, P. J., and Sanders, C. R. (2010) Biochim. Biophys. Acta, 1801, 975–982.
Nadezhdin, K. D., Bocharova, O. V., Bocharov, E. V., and Arseniev, A. S. (2011) Acta Naturae, 3, 69–76.
Nadezhdin, K. D., Bocharova, O. V., Bocharov, E. V., and Arseniev, A. S. (2012) FEBS Lett., 586, 1687–1692.
Istrate, A. N., Tsvetkov, P. O., Mantsyzov, A. B., Kulikova, A. A., Kozin, S. A., Makarov, A. A., and Polshakov, V. I. (2012) Biophys. J., 102, 136–143.
Kozin, S. A., Mezentsev, Y. V., Kulikova, A. A., Indeykina, M. I., Golovin, A. V., Ivanov, A. S., Tsvetkov, P. O., and Makarov, A. A. (2011) Mol. Biosyst., 7, 1053–1055.
Syme, C. D., and Viles, J. H. (2006) Biochim. Biophys. Acta, 1764, 246–256.
Goncharuk, S. A., Goncharuk, M. V., Mayzel, M. L., Lesovoy, D. M., Chupin, V. V., Bocharov, E. V., Arseniev, A. S., and Kirpichnikov, M. P. (2011) Acta Natura, 3, 77–84.
Bocharov, E. V., Mayzel, M. L., Volynsky, P. E., Mineev, K. S., Tkach, E. N., Ermolyuk, Ya. S., Schulga, A. A., Efremov, R. G., and Arseniev, A. S. (2010) Biophys. J., 98, 881–889.
Bocharov, E. V., Mayzel, M. L., Volynsky, P. E., Goncharuk, M. V., Ermolyuk, Ya. S., Schulga, A. A., Artemenko, E. O., Efremov, R. G., and Arseniev, A. S. (2008) J. Biol. Chem., 283, 29385–29395.
Studier, F. W. (2005) Protein Expr. Purif., 41, 207–234.
Kai, L., Roos, C., Haberstock, S., Proverbio, D., Ma, Y., Junge, F., Karbyshev, M., Dotsch, V., and Bernhard, F. (2012) Methods Mol. Biol., 800, 201–225.
Schwarz, D., Junge, F., Durst, F., Frolich, N., Schneider, B., Reckel, S., Sobhanifar, S., Dotsch, V., and Bernhard, F. (2007) Nat. Prot., 2, 2945–2957.
Laemmli, U. K. (1970) Nature, 227, 680–685.
Schagger, H., and von Jagow, G. V. (1987) Anal. Biochem., 166, 368–379.
Bocharova, O. V., Nadezhdin, K. D., Bocharov, E. V., and Arsen’ev, A. S. (2010) Bioorg. Khim., 36, 105–111.
Spirin, A. S., Baranov, V. I., Ryabova, L. A., Ovodov, S. Y., and Alakhov, Y. B. (1988) Science, 242, 1162–1164.
Lundback, A. K., van den Berg, S., Hebert, H., Berglund, H., and Eshaghi, S. (2008) Anal. Biochem., 382, 69–71.
Petrovskaya, L. E., Shulga, A. A., Bocharova, O. V., Ermolyuk, Y. S., Kryukova, E. A., Chupin, V. V., Blommers, M. J., Arseniev, A. S., and Kirpichnikov, M. P. (2010) Biochemistry (Moscow), 75, 881–891.
Bocharova, O. V., Bocharov, E. V., Mineev, K. S., Dubinnyi, M. A., Mishin, A. V., and Arseniev, A. S. (2013) Biochemistry (Moscow). Ser. A: Membr. Cell Biol., 7, 91–99.
Beel, A. J., Mobley, C. K., Kim, H. J., Tian, F., Hadziselimovic, A., Jap, B., Prestegard, J. H., and Sanders, C. R. (2008) Biochemistry, 47, 9428–9446.
Author information
Authors and Affiliations
Corresponding author
Additional information
Published in Russian in Biokhimiya, 2013, Vol. 78, No. 11, pp. 1606–1616.
Rights and permissions
About this article
Cite this article
Bocharova, O.V., Urban, A.S., Nadezhdin, K.D. et al. Bacterial and cell-free production of APP671-726 containing amyloid precursor protein transmembrane and metal-binding domains. Biochemistry Moscow 78, 1263–1271 (2013). https://doi.org/10.1134/S0006297913110060
Received:
Revised:
Published:
Issue Date:
DOI: https://doi.org/10.1134/S0006297913110060