NMR spectroscopic characterization of ribosomal complexes

• Applicant: Professor Dr. Bernd Reif
• Subject Area: Structural Biology
• Term: from 2016 to 2021
• Project identifier: Deutsche Forschungsgemeinschaft (DFG) - Project number 289622162

• Final Report Year: 2022

Final Report Abstract

We have shown that ribosomal complexes can be investigated using MAS solid-state NMR. The achieved sensitivity is sufficient to resolve individual atoms in this asymmetric protein complex. It turned out that reconstitution of labeled proteins in the context of a largely unlabeled background is difficult. It was not possible to prepare ribosome nascent chains of differing length. Also, reconstitution of labeled L23 failed, since it was not possible to grow and harvest cells carrying 50S∆L23.

Publications

• Site-Specific Solid-State NMR Studies of “Trigger Factor” in Complex with the Large Ribosomal Subunit 50S. Angewandte Chemie Int. Edt. Engl. 2015, 54, 4367–4369
  Barbet-Massin, E.; Huang, C.-T.; Daebel, V.; Hsu, S.-T. D.; Reif, B.
  
• Reconstitution of Isotopically Labeled Ribosomal Protein L29 in the 50S Large Ribosomal Subunit for Solution-State and Solid-State NMR. Methods Mol Biol. 2018, 1764, 87-100
  Barbet-Massin, E.; van der Sluis, E.; Musial, J.; Beckmann, R.; Reif, B.