Visualization of a slow, ATP-induced structural transition in the bacterial molecular chaperone DnaK.

The Journal of Biological Chemistry
C D FarrStephan N Witt

Abstract

Recent reports have shown that the binding of ATP to a 70-kDa molecular chaperone induces a rapid global conformational transition from a "high affinity" state to a "low affinity" state, where these states are defined by tight and weak binding to (poly)peptides, respectively. To complete the activity cycle, a chaperone molecule must ultimately return to the high affinity state. In this report, this return to the high affinity state was studied using a chemical cross-linking assay in conjunction with SDS-polyacrylamide gel electrophoresis. The basis for this assay is that in the absence of nucleotide or in the presence of ADP, conditions that stabilize the high affinity state, cross-linking of the Escherichia coli molecular chaperone DnaK yielded two monomeric forms, with apparent molecular masses of 70 kDa (77%) and 90 kDa (23%), whereas cross-linking yielded only the 70-kDa monomeric form in the presence of ATP. This ATP-dependent difference in cross-linking was used to follow the kinetics of the low affinity to high affinity transition under single turnover conditions. The rate of this transition (kobs = 3.4 (+/-0.6) x 10(-4) s-1 at 25 degrees C) is almost identical to the reported rate of ATP hydrolysis (khy = 2.7 (+/-0.7) x...Continue Reading

References

Aug 5, 1978·Journal of Molecular Biology·C M AndersonT A Steitz
Aug 1, 1984·The Journal of Cell Biology·D M SchlossmanJ E Rothman
Jan 15, 1995·The Biochemical Journal·J G Hoggett, G L Kellett
Jan 1, 1993·Annual Review of Biochemistry·J P Hendrick, F U Hartl
Feb 1, 1994·Trends in Biochemical Sciences·R A StuartW Neupert
Apr 1, 1994·European Journal of Biochemistry·N BenaroudjM M Ladjimi
Dec 20, 1993·FEBS Letters·D R PallerosA L Fink
Jan 1, 1993·Annual Review of Cell Biology·C Georgopoulos, W J Welch
Feb 18, 1994·Science·D SchmidP Christen
Nov 15, 1996·Journal of Molecular Biology·H TheyssenJ Reinstein

❮ Previous
Next ❯

Citations

Feb 17, 2001·Journal of Molecular Recognition : JMR·A P Demchenko
Apr 25, 2000·The Journal of Biological Chemistry·M ChevalierS Y Blond
Jun 7, 2011·The Journal of Biological Chemistry·José Ángel Fernández-HigueroArturo Muga
Nov 30, 2004·Biochemical and Biophysical Research Communications·Kazuaki YoshimuneMitsuaki Moriguchi
Feb 16, 2010·Protein Expression and Purification·Wen-I LuoJames A Cowan
Oct 21, 1999·Journal of Structural Biology·J F HainfeldM Barcena
Mar 14, 2000·The Journal of Biological Chemistry·J J Silberg, L E Vickery
Oct 24, 2001·The Journal of Biological Chemistry·Stephanie CarmicleSamuel J Landry
Nov 9, 2005·The Journal of Biological Chemistry·Joanna F SwainLila M Gierasch
Jan 19, 2010·Biochemistry·Stefka G TanevaArturo Muga

❮ Previous
Next ❯

Related Concepts

Related Feeds

ASBMB Publications

The American Society for Biochemistry and Molecular Biology (ASBMB) includes the Journal of Biological Chemistry, Molecular & Cellular Proteomics, and the Journal of Lipid Research. Discover the latest research from ASBMB here.