Structural analysis of GroE chaperonin complexes using chemical cross-linking.

Détails

ID Serval
serval:BIB_4C28886104C2
Type
Article: article d'un périodique ou d'un magazine.
Sous-type
Synthèse (review): revue aussi complète que possible des connaissances sur un sujet, rédigée à partir de l'analyse exhaustive des travaux publiés.
Collection
Publications
Titre
Structural analysis of GroE chaperonin complexes using chemical cross-linking.
Périodique
Methods in Enzymology
Auteur(s)
Azem A., Weiss C., Goloubinoff P.
ISSN
0076-6879 (Print)
ISSN-L
0076-6879
Statut éditorial
Publié
Date de publication
1998
Volume
290
Pages
253-268
Langue
anglais
Résumé
In this chapter, we have shown how chemical cross-linking with a bifunctional reagent, GA, can be used to investigate the structure of large oligomeric complexes such as GroEL14GroES7 and GroEL14(GroES7)2. Cross-linking, followed by denaturing electrophoresis, confirmed the number and arrangement of GroEL and GroES subunits within each individual oligomer, which was previously known from EM analysis. Furthermore, cross-linking permitted a close examination of the effect of regulatory factors, such as nucleotides and free divalent cations, on the molecular structure of GroEL14, GroEL14GroES7, and GroEL14GroES7. Finally, cross-linking analysis permitted characterization and quantitation of various chaperonin heterooligomeric complexes, GroEL14, GroEL14GroES7, and GroEL14GroES7 in solution, under conditions that also supported protein folding and ATP hydrolysis. It was shown that GA does not induce the artifactual association or the dissociation of GroES7 from the chaperonin. On the contrary, chemical cross-linking is an obligatory procedure when the subsequent analysis is carried out using methods that can displace the equilibrium.
Mots-clé
Adenosine Triphosphate/metabolism, Chaperonin 10/chemistry, Chaperonin 60/chemistry, Cross-Linking Reagents/metabolism, Electrophoresis, Polyacrylamide Gel, Escherichia coli/chemistry, Glutaral/metabolism, Microscopy, Electron, Models, Molecular, Protein Conformation
Pubmed
Web of science
Création de la notice
24/01/2008 21:02
Dernière modification de la notice
20/08/2019 15:00
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