Stabilizing Nitroxide Spin Labels for Structural and Conformational Studies of Biomolecules by Maleimide Treatment.
DEER measurement
cell lysates
protein spin labelling
protein structure
stability of NO radical
Journal
Chemistry (Weinheim an der Bergstrasse, Germany)
ISSN: 1521-3765
Titre abrégé: Chemistry
Pays: Germany
ID NLM: 9513783
Informations de publication
Date de publication:
06 Sep 2023
06 Sep 2023
Historique:
received:
28
04
2023
medline:
8
9
2023
pubmed:
24
6
2023
entrez:
24
6
2023
Statut:
ppublish
Résumé
Nitroxide (NO) spin radicals are effective in characterizing structures, interactions and dynamics of biomolecules. The EPR applications in cell lysates or intracellular milieu require stable spin labels, but NO radicals are unstable in such conditions. We showed that the destabilization of NO radicals in cell lysates or even in cells is caused by NADPH/NADH related enzymes, but not by the commonly believed reducing reagents such as GSH. Maleimide stabilizes the NO radicals in the cell lysates by consumption of the NADPH/NADH that are essential for the enzymes involved in destabilizing NO radicals, instead of serving as the solo thiol scavenger. The maleimide treatment retains the crowding properties of the intracellular components and allows to perform long-time EPR measurements of NO labeled biomolecules close to the intracellular conditions. The strategy of maleimide treatment on cell lysates for the EPR applications has been demonstrated on double electron-electron resonance (DEER) measurements on a number of NO labeled protein samples. The method opens a broad application range for the NO labeled biomolecules by EPR in conditions that resemble the intracellular milieu.
Identifiants
pubmed: 37354082
doi: 10.1002/chem.202301350
doi:
Substances chimiques
Spin Labels
0
nitroxyl
GFQ4MMS07W
NAD
0U46U6E8UK
NADP
53-59-8
Maleimides
0
Types de publication
Journal Article
Langues
eng
Sous-ensembles de citation
IM
Pagination
e202301350Subventions
Organisme : Ministry of Science and Technology of China
ID : 2021YFA1600304
Organisme : National Natural Science Foundation of China
ID : 22161142018, 22174074, and 21991081
Informations de copyright
© 2023 Wiley-VCH GmbH.
Références
G. Jeschke, Y. Polyhach, Phys. Chem. Chem. Phys. 2007, 9, 1895-1910;
G. Jeschke, Annu. Rev. Phys. Chem. 2012, 63, 419-446;
D. Goldfarb, Curr. Opin. Struct. Biol. 2022, 75, 102398;
O. Schiemann, T. F. Prisner, Q. Rev. Biophys. 2007, 40, 1-53;
O. Schiemann, C. A. Heubach, D. Abdullin, K. Ackermann, M. Azarkh, E. G. Bagryanskaya, M. Drescher, B. Endeward, J. H. Freed, L. Galazzo, D. Goldfarb, T. Hett, L. E. Hofer, L. F. Ibáñez, E. J. Hustedt, S. Kucher, I. Kuprov, J. E. Lovett, A. Meyer, S. Ruthstein, S. Saxena, S. Stoll, C. R. Timmel, M. Di Valentin, H. S. Mchaourab, T. F. Prisner, B. E. Bode, E. Bordignon, M. Bennati, G. Jeschke, J. Am. Chem. Soc. 2021, 143, 17875-17890;
E. Bordigon, M. A. Seeger, L. Galazzo, G. Meier, FEBS Lett. 2020, 594, 3839-3856.
W. L. Hubbell, D. S. Cafiso, C. Altenbach, Nat. Struct. Biol. 2000, 7, 735-739.
M. Azarkh, V. Singh, O. Okle, I. T. Seemann, D. R. Dietrich, J. S. Hartig, M. Drescher, Nat. Protoc. 2013, 8, 131-147;
D. Nguyen, D. Abdullin, C. A. Heubach, T. Pfaffeneder, A. Nguyen, A. Heine, K. Reuter, F. Diederich, O. Schiemann, G. Klebe, Angew. Chem. Int. Ed. 2021, 60, 23419-23426;
Angew. Chem. 2021, 133, 23607-23615;
A. P. Todd, J. P. Cong, F. Levinthal, C. Levinthal, W. L. Hubell, Proteins Struct. Funct. Genet. 1989, 6, 294-305;
Y. Polyhach, E. Bordignon, R. Tschaggelar, S. Gandra, A. Godt, G. Jeschke, Phys. Chem. Chem. Phys. 2012, 14, 10762-10773;
H. Russell, R. Cura, J. E. Lovett, Front. Mol. Biosci. 2022, 9, 915167;
Q. Cai, A. K. Kusnetzow, W. L. Hubbell, I. S. Haworth, G. P. C. Gacho, N. V. Eps, K. Hideg, E. J. Chambers, P. Z. Qin, Nucleic Acids Res. 2006, 34, 4722-4730.
M. Azarkh, O. Okle, P. Eyring, D. R. Dietrich, M. Drescher, J. Magn. Reson. 2011, 212, 450-454;
M. Teucher, H. Zhang, V. Bader, K. F. Winklhofer, A. J. García-Sáez, A. Rajca, S. Bleicken, E. Bordignon, Sci. Rep. 2019, 9, 13013;
S. Ketter, M. Dajka, O. Rogozhnikova, S. A. Dobrynin, V. M. Tormyshev, E. G. Bagryanskaya, B. Joseph, J. Mag. Reson. Open 2022, 10-11, 100041;
S. Kettter, A. Gopinath, O. Rogozhnikova, D. Trukhin, V. M. Tormyshev, E. G. Bagryanskaya, B. Joseph, Chem. Eur. J. 2021, 27, 2299-2304.
A. Martorana, G. Bellapadrona, A. Feintuch, E. Di Gregorio, S. Aime, D. Goldfarb, J. Am. Chem. Soc. 2014, 136, 13458-13465;
M. Qi, A. Groß, G. Jeschke, A. Godt, M. Drescher, J. Am. Chem. Soc. 2014, 136, 15366-15378;
Y. Yang, F. Yang, Y. J. Gong, J. L. Chen, D. Goldfarb, X. C. Su, Angew. Chem. Int. Ed. 2017, 56, 2914-2918;
Angew. Chem. 2017, 129, 2960-2964.
N. Fleck, C. A. Heubach, T. Hett, F. R. Haege, P. P. Bawol, H. Baltruschat, O. Schiemann, Angew. Chem. Int. Ed. 2020, 59, 9767-9772;
Angew. Chem. 2020, 132, 9854-9859;
Y. Yang, B. B. Pan, X. L. Tan, F. Yang, Y. P. Liu, X. C. Su, D. Goldfarb, J. Phys. Chem. Lett. 2020, 11, 1141-1147.
Y. Polyhach, E. Bordignon, G. Jeschke, Phys. Chem. Chem. Phys. 2011, 13, 2356-2366;
G. Hagelueken, R. Ward, J. H. Naismith, O. Schiemann, Appl. Magn. Reson. 2012, 42, 377-391.
S. Bleicken, T. E. Assafa, H. Zhang, C. Elsner, I. Ritsch, M. Pink, S. Rajca, G. Jeschke, A. Rajca, E. Bordignon, ChemistryOpen 2019, 8, 1057-1065;
G. Karthikeyan, A. Bonucci, G. Casano, G. Gerbaud, S. Abel, V. Thomé, L. Kodjabachian, A. Magalon, B. Guigliarelli, V. Belle, O. Ouari, E. Mileo, Angew. Chem. Int. Ed. 2018, 57, 1366-1370;
Angew. Chem. 2018, 130, 1380-1384;
A. P. Jagtap, I. Krstic, N. C. Kunjir, R. Hänsel, T. F. Prisner, S. T. Sigurdsson, Free Radical Res. 2015, 49, 78-85;
J. T. Paletta, M. Pink, B. Foley, S. Rajca, A. Rajca, Org. Lett. 2012, 14, 5322-5325;
A. Collauto, S. von Bülow, D. B. Gophane, S. Saha, L. S. Stelzl, G. Hummer, S. T. Sigurdsson, T. F. Prisner, Angew. Chem. Int. Ed. 2020, 59, 23025-23029;
Angew. Chem. 2020, 132, 23225-23229.
K. Singewald, M. J. Lawless, S. Saxena, J. Magn. Reson. 2019, 299, 21-27;
M. J. Lawless, A. Shimshi, T. F. Cunningham, M. N. Kinde, P. Tang, S. Saxena, ChemPhysChem 2017, 18, 1653-1660.
Z. Serber, P. Selenko, R. Hänsel, S. Reckel, F. Löhr, J. E. Ferrell, G. Wagner, V. Dötsch, Nat. Protoc. 2006, 1, 2701-2709.
S. Leeb, T. Sörensen, F. Yang, X. Mu, M. Oliveberg, J. Danielsson, Curr. Res. Struct. Biol. 2020, 2, 68-78;
S. L. Speer, C. J. Stewart, L. Sapir, D. Harries, G. J. Pielak, Annu. Rev. Biophys. 2022, 51, 267-300.
I. Martínez-Reyes, N. S. Chandel, Nat. Rev. Cancer 2021, 21, 669-680.
A. Bonucci, O. Ouari, B. Guigliarelli, V. Belle, E. Mileo, ChemBioChem 2020, 21, 451-460;
Z. Hasanbasri, K. Singewald, T. D. Gluth, B. Driesschaert, S. Saxena, J. Phys. Chem. B 2021, 125, 5265-5274;
A. Pierro, A. Bonucci, D. Normanno, M. Ansaldi, E. Pilet, O. Ouari, B. Guigliarelli, E. Etienne, G. Gerbaud, A. Magalon, V. Belle, E Mileo, Chem. Eur. J. 2022, 28, e202202249;
Z. Y. Yang, Y. P. Liu, P. Borbat, J. L. Zweier, J. H. Freed, W. L. Hubbell, J. Am. Chem. Soc. 2012, 134, 9950-9952;
A. Pierro, M. Drescher, Chem. Commun. 2023, 59, 1274-1284.
K. M. McCoy, R. Rogawski, O. Stovicek, A. E. McDermott, J. Magn. Reson. 2019, 303, 115-120;
R. Ghosh, R. Dumarieh, Y. L. Xiao, K. K. Frederick, J. Magn. Reson. 2022, 336, 107150.
C. Y. Cui, B. Li, X. C. Su, ACS Cent. Sci. 2023, 10.1021/acscentsci.3c00385.
V. I. Lushchak, J. Amino Acids 2012, 736837;
C. Hwang, A. J. Sinskey, H. F. Lodish, Science 1992, 257, 1496-1502;
K. Umezawa, M. Yoshida, M. Kamiya, T. Yamasoba, Y. Urano, Nat. Chem. 2017, 9, 279-286.
K. Bedard, K. H. Krause, Physiol. Rev. 2007, 87, 245-313;
W. S. Xiao, R. S. Wang, D. E. Handy, J. Loscalzo, Antioxid. Redox Signaling 2018, 28, 251-272;
B. Alberts, A. Johnson, J. Lewis, D. Morgan, M. Raff, K. Roberts, P. Walter, Molecular Biology of the Cell, 6th Ed., Garland Science, 2014;
W. Ying, Antioxid. Redox Signaling 2008, 10, 179-206.
C. Y. Cui, B. Li, D. Cheng, X. Y. Li, J. L. Chen, Y. T. Chen, X. C. Su, Anal. Chem. 2022, 94, 901-908.
A. Iannone, A. Bini, H. M. Swartz, A. Tomasi, V. Vannini, Biochem. Pharmacol. 1989, 38, 2581-2586.
G. Jeschke, V. Chechik, P. Ionita, A. Godt, H. Zimmermann, J. Banham, C. R. Timmel, D. Hilger, H. Jung, Appl. Magn. Reson. 2006, 30, 473-498.
A. S. Maltsev, A. Grishaev, J. Roche, M. Zasloff, A. Bax, J. Am. Chem. Soc. 2014, 136, 3752-3755.
J. Jee, I. J. L. Byeon, J. M. Louis, A. M. Gronenborn, Proteins Struct. Funct. Bioinf. 2008, 71, 1420-1431.
S. L. Speer, W. W. Zheng, X. Jiang, I. T. Chu, A. J. Guseman, M. L. Liu, G. J. Pielak, C. G. Li, Proc. Natl. Acad. Sci. USA 2021, 118, e2019918118.
Y. Yang, S. N. Chen, F. Yang, X. Y. Li, A. Feintuch, X. C. Su, D. Goldfarb, Proc. Natl. Acad. Sci. USA 2020, 117, 20566-20575.
J. B. Wang, X. B. Zuo, P. Yu, I. J. L. Byeon, J. Jung, X. X. Wang, M. Dyba, S. Seifert, C. D. Schwieters, J. Qin, A. M. Gronenborn, Y. X. Wang, J. Am. Chem. Soc. 2009, 131, 10507-10515.
A. Dalaloyan, A. Martorana, Y. Barak, D. Gataulin, E. Reuveny, A. Howe, M. Elbaum, S. Albeck, T. Unger, V. Frydman, E. H. Abdelkader, G. Otting, D. Goldfarb, ChemPhysChem 2019, 20, 1860-1868.
S. W. Perry, L. G. Epstein, H. A. Gelbard, BioTechniques 1997, 22, 1020-1024.
M. Pannier, S. Veit, A. Godt, G. Jeschke, H. W. Spiess, J. Magn. Reson. 2000, 142, 331-340.