Design of cross-linked RNA/protein complexes for structural studies

Autor: Clément Dégut, Ronald Micura, Carine Tisné, Pierre Barraud, Luc Ponchon, Veronika Schwarz
Přispěvatelé: Laboratoire de cristallographie et RMN biologiques (LCRB - UMR 8015), Université Paris Descartes - Paris 5 (UPD5)-Institut de Chimie du CNRS (INC)-Centre National de la Recherche Scientifique (CNRS), Leopold Franzens Universität Innsbruck - University of Innsbruck, Expression Génétique Microbienne (EGM (UMR_8261 / FRE_3630)), Institut de biologie physico-chimique (IBPC (FR_550)), Centre National de la Recherche Scientifique (CNRS)-Centre National de la Recherche Scientifique (CNRS)-Université Paris Diderot - Paris 7 (UPD7)-Centre National de la Recherche Scientifique (CNRS), Centre National de la Recherche Scientifique (CNRS)-Université Paris Descartes - Paris 5 (UPD5), Institut de biologie physico-chimique (IBPC)
Rok vydání: 2019
Předmět:
0301 basic medicine
Models
Molecular
TRNA modification
Biochemistry
Substrate Specificity
03 medical and health sciences
Bacterial Proteins
RNA
Transfer
[SDV.BBM.BC]Life Sciences [q-bio]/Biochemistry
Molecular Biology/Biochemistry [q-bio.BM]
ComputingMilieux_MISCELLANEOUS
chemistry.chemical_classification
tRNA Methyltransferases
Crystallography
030102 biochemistry & molecular biology
biology
[SDV.BBM.BS]Life Sciences [q-bio]/Biochemistry
Molecular Biology/Structural Biology [q-bio.BM]
Thermus thermophilus
RNA
Substrate (chemistry)
General Medicine
biology.organism_classification
Stem-loop
[SDV.BBM.BC]Life Sciences [q-bio]/Biochemistry
Molecular Biology/Biomolecules [q-bio.BM]
[SDV.BBM.BS]Life Sciences [q-bio]/Biochemistry
Molecular Biology/Biomolecules [q-bio.BM]
RNA
Bacterial
030104 developmental biology
Enzyme
chemistry
Covalent bond
Transfer RNA
Biophysics
Protein Binding
Zdroj: Biochimie
Biochimie, Elsevier, 2019, 164, pp.95-98. ⟨10.1016/j.biochi.2019.03.021⟩
ISSN: 1638-6183
0300-9084
DOI: 10.1016/j.biochi.2019.03.021⟩
Popis: Crystallographic studies of RNA/protein complexes are primordial for the understanding of recognition determinants and catalytic mechanisms in the case of enzymes. However, due to the flexibility and propensity to conformational heterogeneity of RNAs, as well as the mostly electrostatic interactions of RNA/protein complexes, they are difficult to crystallize. We present here a method to trap the two interacting partners in a covalent complex, based on a modified reactive RNA allowing the use of the full range of common crystallogenesis tools. We demonstrate the practicability of our approach with the production of a covalent complex of the Thermus thermophilus m1A58 tRNA modification enzyme, and a modified stem loop mimicking the natural substrate of the enzyme.
Databáze: OpenAIRE
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