Unraveling self-assembly pathways of the 468-kDa proteolytic machine TET2.

Abstract : The spontaneous formation of biological higher-order structures from smaller building blocks, called self-assembly, is a fundamental attribute of life. Although the protein self-assembly is a time-dependent process that occurs at the molecular level, its current understanding originates either from static structures of trapped intermediates or from modeling. Nuclear magnetic resonance (NMR) spectroscopy has the unique ability to monitor structural changes in real time; however, its size limitation and time-resolution constraints remain a challenge when studying the self-assembly of large biological particles. We report the application of methyl-specific isotopic labeling combined with relaxation-optimized NMR spectroscopy to overcome both size- and time-scale limitations. We report for the first time the self-assembly process of a half-megadalton protein complex that was monitored at the structural level, including the characterization of intermediate states, using a mutagenesis-free strategy. NMR was used to obtain individual kinetics data on the different transient intermediates and the formation of final native particle. In addition, complementary time-resolved electron microscopy and native mass spectrometry were used to characterize the low-resolution structures of oligomerization intermediates.
Type de document :
Article dans une revue
Science Advances , American Association for the Advancement of Science (AAAS), 2017, 3 (4), pp.e1601601. 〈10.1126/sciadv.1601601〉
Liste complète des métadonnées

http://hal.univ-grenoble-alpes.fr/hal-01561616
Contributeur : Frank Thomas <>
Soumis le : jeudi 13 juillet 2017 - 10:32:39
Dernière modification le : lundi 19 février 2018 - 14:34:03

Lien texte intégral

Identifiants

Collections

Citation

Pavel Macek, Rime Kerfah, Elisabetta Boeri Erba, Elodie Crublet, Christine Moriscot, et al.. Unraveling self-assembly pathways of the 468-kDa proteolytic machine TET2.. Science Advances , American Association for the Advancement of Science (AAAS), 2017, 3 (4), pp.e1601601. 〈10.1126/sciadv.1601601〉. 〈hal-01561616〉

Partager

Métriques

Consultations de la notice

209