Robust cell-free expression of sub-pathological and pathological huntingtin Exon-1 for NMR studies. General approaches for the isotopic labeling of low-complexity proteins
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The high-resolution structural study of huntingtin exon-1 (HttEx1) has long been hampered by its intrinsic properties. In addition to being prone to aggregate, HttEx1 contains low-complexity regions (LCRs) and is intrinsically disordered, ruling out several standard structural biology approaches. Here, we use a cell-free (CF) protein expression system to robustly and rapidly synthesize (sub-) pathological HttEx1. The open nature of the CF reaction allows the application of different isotopic labeling schemes, making HttEx1 amenable for nuclear magnetic resonance studies. While uniform and selective labeling facilitate the sequential assignment of HttEx1, combining CF expression with nonsense suppression allows the site-specific incorporation of a single labeled residue, making possible the detailed investigation of the LCRs. To optimize CF suppression yields, we analyze the expression and suppression kinetics, revealing that high concentrations of loaded suppressor tRNA have a negative impact on the final reaction yield. The optimized CF protein expression and suppression system is very versatile and well suited to produce challenging proteins with LCRs in order to enable the characterization of their structure and dynamics.
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High-resolution structural studies of purely monomeric species are challenging because these molecules tend to self-associate readily.
“High-resolution structural studies of purely monomeric species are challenging because these molecules tend to selfassociate readily.89,90”
From AlphaFold and Implications for Intrinsically Disordered Proteins (2021) · cites this paper as [89]
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Protein production by auto-induction in high-density shaking cultures
Studier, F. W. (2005) · Protein Expression and Purification · doi:10.1016/j.pep.2005.01.016 · Cited by 5,949 · see citing works
The CCPN data model for NMR spectroscopy: Development of a software pipeline
Vranken, W., Boucher, W., Stevens, T. J., Fogh, R. H., Pajon, A., Llinás, M., Ulrich, E. L., Markley, J. L., Ionides, J., & Laue, E. D. (2005) · Proteins Structure Function and Bioinformatics · doi:10.1002/prot.20449 · Cited by 3,222 · see citing works
Mangiarini, L., Sathasivam, K., Seller, M., Cozens, B., Harper, A., Hetherington, C., Lawton, M., Trottier, Y., Lehrach, H., Davies, S. W., et al. (1996) · Cell · doi:10.1016/s0092-8674(00)81369-0 · Cited by 3,073 · see citing works
Walker, F. O. (2007) · The Lancet · doi:10.1016/s0140-6736(07)60111-1 · Cited by 2,623 · see citing works
Engineering and characterization of a superfolder green fluorescent protein
Pédelacq, J. D., Cabantous, S., Tran, T. H., Terwilliger, T. C., & Waldo, G. S. (2005) · Nature Biotechnology · doi:10.1038/nbt1172 · Cited by 2,295 · see citing works
Wishart, D. S., & Sykes, B. D. (1994) · Journal of Biomolecular NMR · doi:10.1007/bf00175245 · Cited by 2,144 · see citing works
Wishart, D. S., Sykes, B. D., & Richards, F. M. (1992) · Biochemistry · doi:10.1021/bi00121a010 · Cited by 2,057 · see citing works
Wang, J., Choi, J.-M., Holehouse, A., Lee, H. O., Zhang, X., Jahnel, M., Maharana, S., Lemaitre, R., Pozniakovsky, A., Drechsel, D., et al. (2018) · Cell · doi:10.1016/j.cell.2018.06.006 · Cited by 1,876 · see citing works
Scherzinger, E., Lurz, R., Turmaine, M., Mangiarini, L., Hollenbach, B., Hasenbank, R., Bates, G. P., Davies, S. W., Lehrach, H., & Wanker, E. E. (1997) · Cell · doi:10.1016/s0092-8674(00)80514-0 · Cited by 1,291 · cited 1× · see citing works
“The N-terminal domain, preceding the poly-Q tract, is composed of 17 residues (N17) and has been shown to enhance the aggregation of poly-Q tracts in vitro and in vivo [14,15].”
BackgroundA General Method for Site-specific Incorporation of Unnatural Amino Acids into Proteins
Noren, C. J., Anthony-Cahill, S. J., Griffith, M. C., & Schultz, P. G. (1989) · Science · doi:10.1126/science.2649980 · Cited by 1,019 · see citing works
Saudou, F., & Humbert, S. (2016) · Neuron · doi:10.1016/j.neuron.2016.02.003 · Cited by 945 · see citing works
Molecular Mechanisms and Potential Therapeutical Targets in Huntington's Disease
Zuccato, C., Valenza, M., & Cattaneo, E. (2010) · Physiological Reviews · doi:10.1152/physrev.00041.2009 · Cited by 848 · see citing works
Pak, C. W., Kosno, M., Holehouse, A., Padrick, S., Mittal, A., Ali, R., Yunus, A., Liu, D. R., Pappu, R., & Rosen, M. (2016) · Molecules and Cells · doi:10.1016/j.molcel.2016.05.042 · Cited by 747 · see citing works
SUMO Modification of Huntingtin and Huntington's Disease Pathology
Steffan, J. S., Agrawal, N., Pallos, J., Rockabrand, E., Trotman, L. C., Slepko, N., Illes, K., Lukácsovich, T., Zhu, Y., Cattaneo, E. F., et al. (2004) · Science · doi:10.1126/science.1092194 · Cited by 713 · see citing works
Graham, R. K., Deng, Y., Slow, E., Haigh, B., Bissada, N., Lu, G., Pearson, J., Shehadeh, J., Bertram, L. N., Murphy, Z., et al. (2006) · Cell · doi:10.1016/j.cell.2006.04.026 · Cited by 652 · see citing works
Kremer, B., Goldberg, P., Andrew, S. E., Theilmann, J., Telenius, H., Zeisler, J., Squitieri, F., Lin, B., Bassett, A., Almqvist, E., et al. (1994) · New England Journal of Medicine · doi:10.1056/nejm199405193302001 · Cited by 602 · see citing works
Bremer, H., & Dennis, P. P. (2008) · EcoSal Plus · doi:10.1128/ecosal.5.2.3 · Cited by 539 · see citing works
Cell‐free production and stable‐isotope labeling of milligram quantities of proteins
Kigawa, T., Yabuki, T., Yoshida, Y., Tsutsui, M., Ito, Y., Shibata, T., & Yokoyama, S. (1999) · FEBS Letters · doi:10.1016/s0014-5793(98)01620-2 · Cited by 491 · see citing works
Optimal isotope labelling for NMR protein structure determinations
Kainosho, M., Torizawa, T., Iwashita, Y., Terauchi, T., Ono, A. M., & Güntert, P. (2006) · Nature · doi:10.1038/nature04525 · Cited by 485 · see citing works
Polyproline and the “spectroscopic ruler” revisited with single-molecule fluorescence
Schuler, B., Lipman, E. A., Steinbach, P. J., Kumke, M. U., & Eaton, W. A. (2005) · Proceedings of the National Academy of Sciences · doi:10.1073/pnas.0408164102 · Cited by 457 · see citing works
RefDB: A database of uniformly referenced protein chemical shifts
Zhang, H., Neal, S., & Wishart, D. S. (2003) · Journal of Biomolecular NMR · doi:10.1023/a:1022836027055 · Cited by 448 · see citing works
Thakur, A. K., Jayaraman, M., Mishra, R. K., Thakur, M., Chellgren, V. M., Byeon, I. L., Anjum, D. H., Kodali, R. B., Creamer, T. P., Conway, J. F., et al. (2009) · Nature Structural & Molecular Biology · doi:10.1038/nsmb.1570 · Cited by 438 · see citing works
Protein synthesis by pure translation systems
Shimizu, Y., Kanamori, T., & Ueda, T. (2005) · Methods · doi:10.1016/j.ymeth.2005.04.006 · Cited by 413 · see citing works
High-throughput preparation methods of crude extract for robust cell-free protein synthesis
Kwon, Y., & Jewett, M. C. (2015) · Scientific Reports · doi:10.1038/srep08663 · Cited by 382 · see citing works
Preparation of Escherichia coli cell extract for highly productive cell-free protein expression
Kigawa, T., Yabuki, T., Matsuda, N., Matsuda, T., Nakajima, R., Tanaka, A., & Yokoyama, S. (2004) · Journal of Structural and Functional Genomics · doi:10.1023/b:jsfg.0000029204.57846.7d · Cited by 330 · see citing works
A Liquid to Solid Phase Transition Underlying Pathological Huntingtin Exon1 Aggregation
Peskett, T. R., Rau, F., O’Driscoll, J., Patani, R., Lowe, A. R., & Saibil, H. R. (2018) · Molecular Cell · doi:10.1016/j.molcel.2018.04.007 · Cited by 321 · see citing works
Sequence-Specific Random Coil Chemical Shifts of Intrinsically Disordered Proteins
Tamiola, K., Acar, B., & Mulder, F. A. A. (2010) · Journal of the American Chemical Society · doi:10.1021/ja105656t · Cited by 315 · see citing works
Recommendations for the presentation of NMR structures of proteins and nucleic acids
Markley, J. L., Bax, A., Arata, Y., Hilbers, C. W., Kaptein, R., Sykes, B. D., Wright, P. E., & Kurt Wüthrich, A. (1998) · Journal of Molecular Biology · doi:10.1006/jmbi.1998.1852 · Cited by 301 · see citing works
Flanking sequences profoundly alter polyglutamine toxicity in yeast
Duennwald, M. L., Jagadish, S., Muchowski, P. J., & Lindquist, S. L. (2006) · Proceedings of the National Academy of Sciences · doi:10.1073/pnas.0604547103 · Cited by 282 · see citing works
Oligoproline Effects on Polyglutamine Conformation and Aggregation
Bhattacharyya, A., Thakur, A. K., Chellgren, V. M., Thiagarajan, G., Williams, A. D., Chellgren, B. W., Creamer, T. P., & Wetzel, R. B. (2005) · Journal of Molecular Biology · doi:10.1016/j.jmb.2005.10.053 · Cited by 266 · see citing works
Secondary Structure of Huntingtin Amino-Terminal Region
Kim, M. W., Chelliah, Y., Kim, S. W., Otwinowski, Z., & Борисович, Б. И. (2009) · Structure · doi:10.1016/j.str.2009.08.002 · Cited by 257 · see citing works
Klammt, C., Schwarz, D. F., Fendler, K., Haase, W., Dötsch, V., & Bernhard, F. (2005) · FEBS Journal · doi:10.1111/j.1742-4658.2005.05002.x · Cited by 203 · see citing works
The cryo-electron microscopy structure of huntingtin
Guo, Q., Huang, B., Cheng, J., Seefelder, M., Engler, T., Pfeifer, G., Oeckl, P., Otto, M., Moser, F., Maurer, M., et al. (2018) · Nature · doi:10.1038/nature25502 · Cited by 188 · see citing works
Cell-free synthesis and amino acid-selective stable isotope labeling of proteins for NMR analysis
Kigawa, T., Muto, Y., & Yokoyama, S. (1995) · Journal of Biomolecular NMR · doi:10.1007/bf00211776 · Cited by 173 · see citing works
Ishihara, G., Goto, M., Saeki, M., Ito, K., Hori, T., Kigawa, T., Shirouzu, M., & Yokoyama, S. (2005) · Protein Expression and Purification · doi:10.1016/j.pep.2005.01.013 · Cited by 173 · see citing works
Darnell, G. D., Orgel, J. P. R. O., Pahl, R., & Meredith, S. C. (2007) · Journal of Molecular Biology · doi:10.1016/j.jmb.2007.09.023 · Cited by 160 · see citing works
A Simple, Robust, and Low-Cost Method To Produce the PURE Cell-Free System
Lavickova, B., & Maerkl, S. J. (2019) · ACS Synthetic Biology · doi:10.1021/acssynbio.8b00427 · Cited by 159 · see citing works
Solution structure of polyglutamine tracts in GST‐polyglutamine fusion proteins
Masino, L., Kelly, G. P., Leonard, K., Trottier, Y., & Pastore, A. (2002) · FEBS Letters · doi:10.1016/s0014-5793(02)02335-9 · Cited by 148 · see citing works
A linear lattice model for polyglutamine in CAG-expansion diseases
Bennett, M. J., Huey‐Tubman, K. E., Herr, A. B., West, A. P., Ross, S. A., & Björkman, P. J. (2002) · Proceedings of the National Academy of Sciences · doi:10.1073/pnas.182393899 · Cited by 144 · see citing works
At the Interface of Chemical and Biological Synthesis: An Expanded Genetic CodePreprint
Xiao, H., & Schultz, P. G. (2016) · Cold Spring Harbor Perspectives in Biology · doi:10.1101/cshperspect.a023945 · Cited by 139 · see citing works
Large scale active-learning-guided exploration for in vitro protein production optimization
Borkowski, O., Koch, M., Zettor, A., Pandi, A., Batista, A. C., Soudier, P., & Faulon, J. (2020) · Nature Communications · doi:10.1038/s41467-020-15798-5 · Cited by 135 · see citing works
Yabuki, T., Kigawa, T., Dohmae, N., Takio, K., Terada, T., Ito, Y., Laue, E. D., Cooper, J. A., Kainosho, M., & Yokoyama, S. (1998) · Journal of Biomolecular NMR · doi:10.1023/a:1008276001545 · Cited by 130 · see citing works
Critical Role of the Proline-rich Region in Huntingtin for Aggregation and Cytotoxicity in Yeast
Dehay, B., & Bertolotti, A. (2006) · Journal of Biological Chemistry · doi:10.1074/jbc.m605558200 · Cited by 130 · see citing works
Efficient production of isotopically labeled proteins by cell-free synthesis: A practical protocol
Torizawa, T., Shimizu, M., Taoka, M., Miyano, H., & Kainosho, M. (2004) · Journal of Biomolecular NMR · doi:10.1007/s10858-004-3534-2 · Cited by 125 · see citing works
Benn, C., Landles, C., He, L., Strand, A. D., Woodman, B., Sathasivam, K., Li, S.-H., Ghazi‐Noori, S., Hockly, E., Faruque, S. M. N. N., et al. (2005) · Human Molecular Genetics · doi:10.1093/hmg/ddi340 · Cited by 123 · see citing works
Using NMR chemical shifts to calculate the propensity for structural order and disorder in proteins
Tamiola, K., & Mulder, F. A. A. (2012) · Biochemical Society Transactions · doi:10.1042/bst20120171 · Cited by 123 · see citing works
Preparation and evaluation of acylated tRNAs
Walker, S. E., & Fredrick, K. (2008) · Methods · doi:10.1016/j.ymeth.2007.09.003 · Cited by 107 · see citing works
Dopp, B. J. L., Tamiev, D., & Reuel, N. F. (2018) · Biotechnology Advances · doi:10.1016/j.biotechadv.2018.12.006 · Cited by 107 · see citing works
Multiple‐Site Labeling of Proteins with Unnatural Amino Acids
Loscha, K. V., Herlt, A. J., Qi, R., Huber, T., Ozawa, K., & Otting, G. (2012) · Angewandte Chemie International Edition · doi:10.1002/anie.201108275 · Cited by 104 · see citing works
Quantification of interlaboratory cell-free protein synthesis variability
Cole, S. D., Beabout, K., Turner, K. B., Smith, Z. K., Funk, V. L., Harbaugh, S. V., Liem, A. T., Roth, P. A., Geier, B. A., Emanuel, P. A., et al. (2019) · ACS Synthetic Biology · doi:10.1021/acssynbio.9b00178 · Cited by 103 · see citing works
Mateos, B., Conrad‐Billroth, C., Schiavina, M., Beier, A., Kontaxis, G., Konrat, R., Felli, I. C., & Pierattelli, R. (2019) · Journal of Molecular Biology · doi:10.1016/j.jmb.2019.11.015 · Cited by 100 · see citing works
Side chain to main chain hydrogen bonds stabilize a polyglutamine helix in a transcription factor
Escobedo, A., Topal, B., Kunze, M. B. A., Aranda, J., Chiesa, G., Mungianu, D., Bernardo‐Seisdedos, G., Eftekharzadeh, B., Gairí, M., Pierattelli, R., et al. (2019) · Nature Communications · doi:10.1038/s41467-019-09923-2 · Cited by 99 · see citing works
Disentangling the complexity of low complexity proteins
Mier, P., Paladin, L., Tamana, S., Petrosian, S., Hajdu-Soltész, B., Urbanek, A., Gruca, A., Plewczyński, D., Grynberg, M., Bernadó, P., et al. (2019) · Briefings in Bioinformatics · doi:10.1093/bib/bbz007 · Cited by 97 · see citing works
Self-citationOzawa, K., Headlam, M. J., Schaeffer, P. M., Henderson, B. R., Dixon, N. E., & Otting, G. (2004) · European Journal of Biochemistry · doi:10.1111/j.1432-1033.2004.04346.x · Cited by 89 · see citing works
Monsellier, É., Redeker, V., Ruiz-Arlandis, G., Bousset, L., & Melki, R. (2014) · Journal of Biological Chemistry · doi:10.1074/jbc.m114.603332 · Cited by 85 · see citing works
Cell-Free Protein Synthesis for Analysis by NMR Spectroscopy
Apponyi, M. A., Ozawa, K., Dixon, N. E., & Otting, G. (2008) · Methods in molecular biology · doi:10.1007/978-1-60327-058-8_16 · Cited by 82 · see citing works
Cell‐Free Transcription/Translation from PCR‐Amplified DNA for High‐Throughput NMR Studies
Wu, P., Ozawa, K., Lim, S. P., Vasudevan, S. G., Dixon, N. E., & Otting, G. (2007) · Angewandte Chemie International Edition · doi:10.1002/anie.200605237 · Cited by 81 · see citing works
The E. coli S30 lysate proteome: A prototype for cell-free protein production
Foshag, D., Henrich, E., Hiller, E., Schäfer, M., Kerger, C., Burger‐Kentischer, A., Díaz‐Moreno, I., García‐Mauriño, S. M., Dötsch, V., Rupp, S., et al. (2017) · New Biotechnology · doi:10.1016/j.nbt.2017.09.005 · Cited by 79 · see citing works
Shen, K., Calamini, B., Fauerbach, J. A., Ma, B., Shahmoradian, S. H., Lachapel, I. L. S., Chiu, W., Lo, D. C., & Frydman, J. (2016) · eLife · doi:10.7554/elife.18065 · Cited by 77 · see citing works
Jorda, J., & Kajava, A. V. (2010) · Advances in protein chemistry and structural biology · doi:10.1016/s1876-1623(10)79002-7 · Cited by 75 · see citing works
Site-specific isotopic labeling of proteins for NMR studies
Ellman, J. A., Volkman, B. F., Mendel, D., Schulz, P. G., & Wemmer, D. E. (1992) · Journal of the American Chemical Society · doi:10.1021/ja00046a080 · Cited by 75 · see citing works
Propensity for cis‐Proline Formation in Unfolded Proteins
Alderson, T. R., Lee, J. H., Charlier, C., Ying, J., & Bax, A. (2017) · ChemBioChem · doi:10.1002/cbic.201700548 · Cited by 75 · see citing works
Isas, J. M., Langen, R., & Siemer, A. B. (2015) · Biochemistry · doi:10.1021/acs.biochem.5b00281 · Cited by 71 · see citing works
Structure and Dynamics of the Huntingtin Exon-1 N-Terminus: A Solution NMR Perspective
Baias, M., Smith, P. E. S., Shen, K., Joachimiak, Ł. A., Żerko, S., Koźmiński, W., Frydman, J., & Frydman, L. (2017) · Journal of the American Chemical Society · doi:10.1021/jacs.6b10893 · Cited by 67 · see citing works
15N‐Labelled proteins by cell‐free protein synthesis
Ozawa, K., Wu, P., Dixon, N. E., & Otting, G. (2006) · FEBS Journal · doi:10.1111/j.1742-4658.2006.05433.x · Cited by 66 · see citing works
Constraints and consequences of the emergence of amino acid repeats in eukaryotic proteins
Chavali, S., Chavali, P. L., Chalancon, G., de Groot, N. S., Gemayel, R., Latysheva, N. S., Ing‐Simmons, E., Verstrepen, K. J., Santhanam, B., & Babu, M. M. (2017) · Nature Structural & Molecular Biology · doi:10.1038/nsmb.3441 · Cited by 66 · see citing works
Newcombe, E. A., Ruff, K. M., Sethi, A., Ormsby, A. R., Ramdzan, Y. M., Fox, A., Purcell, A. W., Gooley, P. R., Pappu, R. V., & Hatters, D. M. (2018) · Journal of Molecular Biology · doi:10.1016/j.jmb.2018.03.031 · Cited by 66 · see citing works
Urbanek, A., Popovic, M., Morató, A., Estaña, A., Elena-Real, C. A., Mier, P., Fournet, A., Allemand, F., Delbecq, S., Andrade-Navarro, M. A., et al. (2020) · Structure · doi:10.1016/j.str.2020.04.008 · Cited by 66 · see citing works
Self-citationDarnell, G. D., Derryberry, J., Kurutz, J. W., & Meredith, S. C. (2009) · Biophysical Journal · doi:10.1016/j.bpj.2009.07.062 · Cited by 60 · see citing works
Sahoo, B., Arduini, I., Drombosky, K. W., Kodali, R. B., Sanders, L. H., Greenamyre, J. T., & Wetzel, R. B. (2016) · PLoS ONE · doi:10.1371/journal.pone.0155747 · Cited by 59 · see citing works
Amino acid homorepeats in proteins
Chavali, S., Singh, A., Santhanam, B., & Babu, M. (2020) · Nature Reviews Chemistry · doi:10.1038/s41570-020-0204-1 · Cited by 58 · see citing works
A practical method for cell-free protein synthesis to avoid stable isotope scrambling and dilution
Yokoyama, J., Matsuda, T., Koshiba, S., Tochio, N., & Kigawa, T. (2011) · Analytical Biochemistry · doi:10.1016/j.ab.2011.01.017 · Cited by 55 · see citing works
Nazarenko, I. A., Harrington, K. M., & Uhlenbeck, O. C. (1994) · The EMBO Journal · doi:10.1002/j.1460-2075.1994.tb06531.x · Cited by 52 · see citing works
Fei, J., Wang, J., Sternberg, S. H., MacDougall, D., Elvekrog, M., Pulukkunat, D. K., Englander, M. T., & Gonzalez, R. L. (2010) · Methods in enzymology on CD-ROM/Methods in enzymology · doi:10.1016/s0076-6879(10)72008-5 · Cited by 52 · see citing works
Trinucleotide Repeats: A Structural Perspective
Almeida, B., Fernandes, S., Abreu, I. A., & Macedo‐Ribeiro, S. (2013) · Frontiers in Neurology · doi:10.3389/fneur.2013.00076 · Cited by 52 · see citing works
The folding equilibrium of huntingtin exon 1 monomer depends on its polyglutamine tract
Bravo-Arredondo, J. M., Kegulian, N. C., Schmidt, T. H., PANDEY, N. K., Situ, A. J., Ulmer, T. S., & Langen, R. (2018) · Journal of Biological Chemistry · doi:10.1074/jbc.ra118.004808 · Cited by 48 · see citing works
(2002) · Springer eBooks · doi:10.1007/978-3-642-59379-6 · Cited by 47 · see citing works
Process optimization for scalable E. coli extract preparation for cell-free protein synthesis
Dopp, J. L., & Reuel, N. F. (2018) · Biochemical Engineering Journal · doi:10.1016/j.bej.2018.06.021 · Cited by 45 · see citing works
Rapid prototyping of proteins: Mail order gene fragments to assayable proteins within 24 hours
Dopp, J. L., Rothstein, S. M., Mansell, T. J., & Reuel, N. F. (2018) · Biotechnology and Bioengineering · doi:10.1002/bit.26912 · Cited by 45 · see citing works
Selective Methyl Labeling of Eukaryotic Membrane Proteins Using Cell-Free Expression
Linser, R., Gelev, V., Hagn, F., Arthanari, H., Hyberts, S. G., & Wagner, G. (2014) · Journal of the American Chemical Society · doi:10.1021/ja504791j · Cited by 42 · see citing works
Artificial Environments for the Co-Translational Stabilization of Cell-Free Expressed Proteins
Kai, L., Dötsch, V., Kaldenhoff, R., & Bernhard, F. (2013) · PLoS ONE · doi:10.1371/journal.pone.0056637 · Cited by 41 · see citing works
Pleiss, J. A., & Uhlenbeck, O. C. (2001) · Journal of Molecular Biology · doi:10.1006/jmbi.2001.4612 · Cited by 41 · see citing works
Urbanek, A., Morató, A., Allemand, F., Delaforge, E., Fournet, A., Popovic, M., Delbecq, S., Sibille, N., & Bernadó, P. (2018) · Angewandte Chemie International Edition · doi:10.1002/anie.201711530 · Cited by 39 · see citing works
Self-citationCell-free synthesis of 15 N-labeled proteins for NMR studies
Ozawa, K., Dixon, N. E., & Otting, G. (2005) · IUBMB Life · doi:10.1080/15216540500217859 · Cited by 36 · see citing works
Evidence of the Reduced Abundance of Proline cis Conformation in Protein Poly Proline Tracts
Urbanek, A., Popovic, M., Elena‐Real, C. A., Morató, A., Estaña, A., Fournet, A., Allemand, F., Gil, A. M., Cativiela, C., Cortés, J., et al. (2020) · Journal of the American Chemical Society · doi:10.1021/jacs.0c02263 · Cited by 36 · see citing works
Self-citationEfficient Isotope Editing of Proteins for Site-Directed Vibrational Spectroscopy
Peuker, S., Andersson, H., Gustavsson, E., Maiti, K. S., Kania, R., Karim, A., Niebling, S., Pedersen, A., Erdélyi, M., & Westenhoff, S. (2016) · Journal of the American Chemical Society · doi:10.1021/jacs.5b12680 · Cited by 35 · see citing works
Proline Fingerprint in Intrinsically Disordered Proteins
Murrali, M. G., Piai, A., Bermel, W., Felli, I. C., & Pierattelli, R. (2018) · ChemBioChem · doi:10.1002/cbic.201800172 · Cited by 34 · see citing works
Wu, Y., Wang, Z., Qiao, X., Li, J., Shu, X., & Qi, H. (2020) · Frontiers in Bioengineering and Biotechnology · doi:10.3389/fbioe.2020.00863 · Cited by 32 · see citing works
Protein labeling strategies for liquid-state NMR spectroscopy using cell-free synthesis
Hoffmann, B., Löhr, F., Laguerre, A., Bernhard, F., & Dötsch, V. (2017) · Progress in Nuclear Magnetic Resonance Spectroscopy · doi:10.1016/j.pnmrs.2017.11.004 · Cited by 30 · see citing works
An economical method for producing stable-isotope labeled proteins by the E. coli cell-free system
Yokoyama, J., Matsuda, T., Koshiba, S., & Kigawa, T. (2010) · Journal of Biomolecular NMR · doi:10.1007/s10858-010-9455-3 · Cited by 20 · see citing works
Urbanek, A., Elena‐Real, C. A., Popovic, M., Morató, A., Fournet, A., Allemand, F., Delbecq, S., Sibille, N., & Bernadó, P. (2019) · ChemBioChem · doi:10.1002/cbic.201900583 · Cited by 12 · see citing works
Self-citationLimiting factors of the translation machinery
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Escobedo, A., Chiesa, G., & Salvatella, X. (2020) · Methods in molecular biology · doi:10.1007/978-1-0716-0524-0_10 · Cited by 4 · see citing works
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The structure of pathogenic huntingtin exon 1 defines the bases of its aggregation propensity
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Elena‐Real, C. A., Urbanek, A., Lund, X. L., Morató, A., Sagar, A., Fournet, A., Estaña, A., Bellande, T., Allemand, F., Cortés, J., et al. (2023) · Structure · doi:10.1016/j.str.2023.04.003 · Cited by 10 · see citing works
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Self-citationEnabling and optimizing cell-free systems for synthetic biology applications
Batista, A. C. (2022) · theses.fr (ABES) · doi:10.70675/60aa7569zfd16z48ecz93b4z907df93453cf
- Authors
- Morató, A., Elena-Real, C. A., Popovic, M., Fournet, A., Zhang, K., Allemand, F., Sibille, N., Urbanek, A., & Bernadó, P.
- Affiliations
- Centre National de la Recherche Scientifique (FR)
- Centre de Biochimie Structurale (CBS), INSERM, CNRS and Université de Montpellier. 29 rue de Navacelles, 34090 Montpellier, France
- Corresponding author
- Urbanek, Annika
- Bernadó, Pau
- ORCID
- Venue
- Biomolecules
- Publisher
- MDPI AG
- Published
- 2020-10-19
- Volume
- 10
- Issue
- 10
- Pages
- 1458
- ISSN
- 2218-273X (ISSN-L)
- PMC
- PMC7603387
- OpenAlex
- W3093375775
- Cited by
- 12
- References
- 93
- Citations by year
- 2025: 1 · 2024: 1 · 2023: 4 · 2022: 4 · 2021: 2
- FWCI
- 0.95
- Type
- journal article
- Language
- en
- Open access
- gold
- doi.org
- Licence
- Locations
- Biomolecules (publishedVersion, cc-by)
- PubMed Central (publishedVersion, cc-by)
- PubMed (publishedVersion)
- HAL (submittedVersion, other-oa)
- DOAJ (DOAJ: Directory of Open Access Journals) (submittedVersion, cc-by-sa)
- Funders
- Agence Nationale de la Recherche (ANR-10-LABX-12-01, ANR-17-CE11-0022-01, ANR-10-INBS-04-01, ANR-10-INBS-05)
- H2020 European Research Council (648030)
- Topics
- Physical Sciences › Chemistry › Spectroscopy › Advanced NMR Techniques and Applications
- Life Sciences › Neuroscience › Cellular and Molecular Neuroscience › Genetic Neurodegenerative Diseases
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › RNA and protein synthesis mechanisms
- Keywords
- huntingtin exon 1, cell-free expression, low complexity regions, intrinsically disordered proteins, isotopic labeling, NMR spectroscopy, nonsense suppression, suppressor tRNA, protein aggregation, protein dynamics, site-specific labeling
- MeSH terms
- Huntingtin Protein, Protein Domains, Cell-Free System, Exons, Humans, Huntington Disease, Isotope Labeling, Kinetics, Protein Processing, Post-Translational, Nuclear Magnetic Resonance, Biomolecular, Protein Aggregation, Pathological
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