SpringerLink
Log in

Preparation, Crystallization, and Preliminary X-Ray Diffraction Study of Mutant Carboxypeptidase T Bearing the Primary Specificity Pocket and the Active-Site Loop of Carboxypeptidase B

  • STRUCTURE OF MACROMOLECULAR COMPOUNDS
  • Published:
Crystallography Reports Aims and scope Submit manuscript

Abstract

Crystals of mutant carboxypeptidase T from Thermoactinomyces vulgaris (CPT11QG) with amino-acid substitutions G215S, Q249G, A251G, T257A, D260G, T262D, and L254I and with the insertion ins253T were grown in microgravity by the capillary counter-diffusion method. The crystals belong to sp. gr. P31, which differs from the space group of the wild-type enzyme (P6322). The X-ray diffraction data set was collected from the crystals at the SPring-8 synchrotron facility (Japan) and is suitable for crystal structure determination at 2.45 Å resolution.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Subscribe and save

Springer+ Basic
EUR 32.99 /Month
  • Get 10 units per month
  • Download Article/Chapter or Ebook
  • 1 Unit = 1 Article or 1 Chapter
  • Cancel anytime
Subscribe now

Buy Now

Price includes VAT (United Kingdom)

Instant access to the full article PDF.

Institutional subscriptions

Similar content being viewed by others

REFERENCES

  1. J. G. Perez-Silva, J. Espanol, G. Velasco, and V. Quesada, Nucleic Acids Res. 44 (1), D351 (2016).

    Article  Google Scholar 

  2. B. Turk, D. Turk, and V. Turk, EMBO J. 31 (7), 1630 (2012).

    Article  Google Scholar 

  3. B. Turk, Nat. Rev. Drug Discovery 5 (9), 785 (2006).

    Article  Google Scholar 

  4. A. Teplyakov, K. Polakov, G. Obmolova, et al., Eur. J. Biochem. 208 (2), 281 (1992).

    Article  Google Scholar 

  5. M. Edge, S. Forder, J. Hennam, et al., Protein Eng. 11 (12), 1229 (1998).

    Article  Google Scholar 

  6. M. C. Garcia-Guerrero, J. Garsia-Prado, E. Beringuer, et al., Proc. Natl. Acad. Sci. U.S.A. 115 (17), E3932 (2018).

    Article  Google Scholar 

  7. V. Akparov, V. Timofeev, I. Khaliullin, et al., J. Biomol. Struct. Dyn. 36 (4), 956 (2018).

    Article  Google Scholar 

  8. A. M. Grishin, V. K. Akparov, and G. G. Chestukhina, Protein Eng. Des. Sel. 21 (9), 545 (2008).

    Article  Google Scholar 

  9. V. Kh. Akparov, A. M. Grishin, M. P. Yusupova, et al., Biokhimiya 72 (4), 515 (2007).

    Google Scholar 

  10. V. Akparov, V. Timofeev, I. Kuranova, and T. Rakitina, Acta Crystallogr. F 74 (10), 638 (2018).

    Article  Google Scholar 

  11. A. M. Grishin, V. Kh. Akparov, and G. G. Chestukhina, Biokhimiya 73 (10), 1422 (2008).

    Google Scholar 

  12. Novagen, pET System Manual TB055, 7th. ed. (Novagen, Madison, WI, 1997).

  13. L. Trachuk, A. Letarov, I. Kudelina, et al., Protein Exper. Purif. 40 (1), 51 (2005).

    Article  Google Scholar 

  14. L. B. Bazzone. T. J. Cueni and J. F. Riordan, Anal. Biochem. 107 (2), 341 (1980).

    Article  Google Scholar 

  15. M. M. Bradford, Anal. Biochem. 72, 248 (1976).

    Article  Google Scholar 

  16. U. K. Laemmli, Nature 227 (5259), 680 (1970).

    Article  ADS  Google Scholar 

  17. S. Takahashi, T. Tsurumura, K. Aritake, et al., Acta Crystallogr. F 66 (7), 846 (2010).

    Article  Google Scholar 

  18. I. P. Kuranova, E. A. Smirnova, Yu. A. Abramchik, et al., Crystallogr. Rep. 56 (5), 884 (2011).

    Article  ADS  Google Scholar 

  19. T. Battye, L. Kontogiannes, O. Johnson, and H. Powel, Acta Crystallogr. D 67 (4), 271 (2011).

    Article  Google Scholar 

  20. B. W. Mattews, J. Mol. Biol. 33, 491 (1968).

    Article  Google Scholar 

  21. M. D. Winn, C. C. Ballard, K. D. Kowtan, et al., Acta Crystallogr. D 67 (4), 235 (2011).

    Article  Google Scholar 

  22. V. K. Akparov, V. I. Timofeev, G. E. Konstantinova, et al., PLOS One 14 (12), 1 (2019).

    Article  Google Scholar 

  23. V. Kh. Akparov, V. I. Timofeev, I. G. Khaliullin, et al., J. Biomol. Struct. Dyn. 36 (15), 3958 (2018).

    Article  Google Scholar 

  24. V. Kh. Akparov, V. I. Timofeev, I. G. Khaliullin, et al., Biokhimiya 84 (1), 53 (2019).

    Google Scholar 

  25. V. Kh. Akparov, A. M. Grishin, V. I. Timofeev, and I. P. Kuranova, Crystallogr. Rep. 55 (5), 802 (2010).

    Article  ADS  Google Scholar 

Download references

Funding

This study was financially supported by the Russian Foundation for Basic Research (project no. 19-04-00220; protein isolation and purification), the Federal Space Program for 2016−2025 (International Space Station Research and Development Program (Nauka); crystal growth in microgravity), and the Ministry of Science and Higher Education of the Russian Federation within the framework of the state assignment for the Federal Scientific Research Centre “Crystallography and Photonics” of the Russian Academy of Sciences (X-ray data collection and processing).

Author information

Authors and Affiliations

  1. National Research Center “Kurchatov Institute”, 123098, Moscow, Russia

    V. Kh. Akparov, G. E. Konstantinova, V. I. Timofeev & I. P. Kuranova

  2. Shubnikov Institute of Crystallography of the Federal Scientific Research Centre “Crystallography and Photonics,” Russian Academy of Sciences, 119333, Moscow, Russia

    V. I. Timofeev & I. P. Kuranova

  3. Moscow Institute of Physics and Technology, 141701, Dolgoprudnyi, Moscow oblast, Russia

    I. G. Khaliullin

Authors
  1. V. Kh. Akparov
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. G. E. Konstantinova
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. V. I. Timofeev
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. I. P. Kuranova
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. I. G. Khaliullin
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding authors

Correspondence to V. Kh. Akparov or V. I. Timofeev.

Additional information

Translated by T. Safonova

Rights and permissions

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Akparov, V.K., Konstantinova, G.E., Timofeev, V.I. et al. Preparation, Crystallization, and Preliminary X-Ray Diffraction Study of Mutant Carboxypeptidase T Bearing the Primary Specificity Pocket and the Active-Site Loop of Carboxypeptidase B. Crystallogr. Rep. 65, 900–902 (2020). https://doi.org/10.1134/S106377452006005X

Download citation

  • Received:

  • Revised:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1134/S106377452006005X

Search

Navigation