Amide hydrogen exchange reveals conformational changes in hsp70 chaperones important for allosteric regulation

The Journal of Biological Chemistry
Wolfgang RistM P Mayer

Abstract

Hsp70 chaperones assist protein folding processes by a nucleotide-driven cycle of substrate binding and release. Although structural information is available for the isolated nucleotide-binding (NBD) and substrate-binding domains (SBD) in the high affinity conformation, the low affinity conformations and the conformational changes associated with mutual allosteric regulation remained largely enigmatic. By using amide hydrogen exchange in combination with mass spectrometry, we analyzed the Escherichia coli Hsp70 homologue DnaK as full-length protein and its individual domains in the nucleotide-free and ATP-bound conformation. We found a surprising degree of flexibility in both domains. The comparison of the full-length protein with the isolated domains demonstrates a mutual stabilization of both domains. This protection from solvent was most pronounced and in addition was nucleotide-dependent in the lowerbeta-sheet of the SBD and the loop that connects the last beta-strand with helix alphaA. Interestingly, the linker region, which connects NBD and SBD and which is close to the protected loop in the SBD, is solvent-exposed in the absence of nucleotide and completely protected from hydrogen exchange in the presence of ATP. Peptide...Continue Reading

References

Jun 1, 1991·Proceedings of the National Academy of Sciences of the United States of America·K M FlahertyK C Holmes
Jan 1, 1966·Advances in Protein Chemistry·A Hvidt, S O Nielsen
Jul 5, 1996·The Journal of Biological Chemistry·M C O'BrienD B McKay
Nov 15, 1996·Journal of Molecular Biology·H TheyssenJ Reinstein
Dec 8, 1998·Biochemistry·S V Slepenkov, S N Witt
Jan 8, 1999·Journal of the American Society for Mass Spectrometry·Z Zhang, A G Marshall
Feb 27, 1999·Protein Science : a Publication of the Protein Society·E B BertelsenF W Dahlquist
May 13, 1999·Proceedings of the National Academy of Sciences of the United States of America·T LaufenB Bukau
Jul 6, 2000·Nature Structural Biology·M P MayerB Bukau
Nov 15, 2000·Biological Chemistry·M P MayerB Bukau
Mar 9, 2002·Science·F Ulrich Hartl, Manajit Hayer-Hartl
Jul 3, 2004·Proceedings of the National Academy of Sciences of the United States of America·Yongbo Zhang, Erik R P Zuiderweg
Feb 4, 2005·Protein Science : a Publication of the Protein Society·Wolfgang RistMatthias P Mayer
Mar 17, 2005·Cellular and Molecular Life Sciences : CMLS·M P Mayer, B Bukau
May 7, 2005·Journal of Molecular Biology·Matthew RevingtonErik R P Zuiderweg
Jun 17, 2005·Analytical Biochemistry·Wolfgang RistThomas J D Jørgensen
Nov 26, 2005·Molecular Cell·Jianwen JiangRui Sousa
Feb 4, 2006·Molecular Cell·Markus VogelMatthias P Mayer

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Citations

Jan 8, 2009·Molecular Genetics and Genomics : MGG·Bhag Singh, Radhey S Gupta
Sep 10, 2013·Trends in Biochemical Sciences·Matthias P Mayer
Oct 20, 2009·Nature Structural & Molecular Biology·Shinji TsutsumiLen Neckers
Feb 1, 2011·Nature Structural & Molecular Biology·Rainer SchlechtMatthias P Mayer
May 28, 2013·Nature Structural & Molecular Biology·Ruifeng QiQinglian Liu
Aug 2, 2007·Photochemical & Photobiological Sciences : Official Journal of the European Photochemistry Association and the European Society for Photobiology·Koji NakayamaTetsuro Majima
Aug 2, 2012·Molecular BioSystems·Anne T GilliesJason E Gestwicki
Oct 25, 2008·Proceedings of the National Academy of Sciences of the United States of America·Claes AndréassonBernd Bukau
May 15, 2009·Proceedings of the National Academy of Sciences of the United States of America·Eric B BertelsenErik R P Zuiderweg
Feb 9, 2012·Proceedings of the National Academy of Sciences of the United States of America·Shinji TsutsumiLeonard M Neckers
Nov 19, 2009·The Journal of Biological Chemistry·Jocelyne FiauxElke Deuerling
May 6, 2011·The Journal of Biological Chemistry·Goran StjepanovicIrmgard Sinning
Jul 20, 2011·The Journal of Biological Chemistry·Robert G SmockLila M Gierasch
Sep 2, 2006·Acta Crystallographica. Section F, Structural Biology and Crystallization Communications·Liam Worrall, Malcolm D Walkinshaw
May 11, 2010·Biochemistry and Cell Biology = Biochimie Et Biologie Cellulaire·Jason C Young
Jan 10, 2013·PLoS Computational Biology·Federica ChiapporiGiulia Morra
Nov 3, 2012·Biochimica Et Biophysica Acta·Sheila S Jaswal
Dec 2, 2009·Proceedings of the National Academy of Sciences of the United States of America·Gunes BozkurtIrmgard Sinning
Jul 19, 2011·Journal of Molecular Biology·Divya Prasanna KumarQinglian Liu
Dec 19, 2009·Journal of Magnetic Resonance·Akash BhattacharyaErik R P Zuiderweg
Apr 14, 2009·Journal of Molecular Biology·Akash BhattacharyaErik R P Zuiderweg
Oct 27, 2007·Protein Science : a Publication of the Protein Society·Sheila S Jaswal, Andrew D Miranker
Oct 10, 2006·Traffic·Rui Sousa, Eileen M Lafer
Feb 23, 2012·The EMBO Journal·Chung-Tien LeeMatthias P Mayer
Sep 25, 2010·Molecular Systems Biology·Robert G SmockLila M Gierasch
Jun 4, 2015·Proceedings of the National Academy of Sciences of the United States of America·Anastasia Zhuravleva, Lila M Gierasch
Nov 6, 2013·Journal of Molecular Biology·Chung-Tien LeeMatthias P Mayer
Jun 29, 2011·Journal of Molecular Biology·Aikaterini RousakiErik R P Zuiderweg
Feb 16, 2010·Protein Expression and Purification·Wen-I LuoJames A Cowan
Oct 15, 2013·Autophagy·François StricherSylviane Muller
Nov 6, 2012·Molecular Cell·Roman KitykMatthias P Mayer
Nov 13, 2007·Molecular Cell·Jianwen JiangRui Sousa
Aug 14, 2010·Molecular Cell·Matthias P Mayer

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