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

Details

Serval ID
serval:BIB_4C28886104C2
Type
Article: article from journal or magazin.
Publication sub-type
Review (review): journal as complete as possible of one specific subject, written based on exhaustive analyses from published work.
Collection
Publications
Title
Structural analysis of GroE chaperonin complexes using chemical cross-linking.
Journal
Methods in Enzymology
Author(s)
Azem A., Weiss C., Goloubinoff P.
ISSN
0076-6879 (Print)
ISSN-L
0076-6879
Publication state
Published
Issued date
1998
Volume
290
Pages
253-268
Language
english
Abstract
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.
Keywords
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
Create date
24/01/2008 20:02
Last modification date
20/08/2019 14:00
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