title: Combining crystallography and EPR: crystal and solution structures of the multidomain cochaperone DnaJ.
abstract: Hsp70 chaperones assist in a large variety of protein-folding processes in the cell. Crucial for these activities is the regulation of Hsp70 by Hsp40 cochaperones. DnaJ, the bacterial homologue of Hsp40, stimulates ATP hydrolysis by DnaK (Hsp70) and thus mediates capture of substrate protein, but is also known to possess chaperone activity of its own. The first structure of a complete functional dimeric DnaJ was determined and the mobility of its individual domains in solution was investigated. Crystal structures of the complete molecular cochaperone DnaJ from Thermus thermophilus comprising the J, GF and C-terminal domains and of the J and GF domains alone showed an ordered GF domain interacting with the J domain. Structure-based EPR spin-labelling studies as well as cross-linking results showed the existence of multiple states of DnaJ in solution with different arrangements of the various domains, which has implications for the function of DnaJ.
authors.affiliation: MPI for Medical Research, Heidelberg, Germany. thomas.barends@mpimf-heidelberg.mpg.de
authors.forename: Thomas R M
authors.initials: TR
authors.lastname: Barends
authors.forename: Richard W W
authors.initials: RW
authors.lastname: Brosi
authors.forename: Andrea
authors.initials: A
authors.lastname: Steinmetz
authors.forename: Anna
authors.initials: A
authors.lastname: Scherer
authors.forename: Elisabeth
authors.initials: E
authors.lastname: Hartmann
authors.forename: Jessica
authors.initials: J
authors.lastname: Eschenbach
authors.forename: Thorsten
authors.initials: T
authors.lastname: Lorenz
authors.forename: Ralf
authors.initials: R
authors.lastname: Seidel
authors.forename: Robert L
authors.initials: RL
authors.lastname: Shoeman
authors.forename: Sabine
authors.initials: S
authors.lastname: Zimmermann
authors.forename: Robert
authors.initials: R
authors.lastname: Bittl
authors.forename: Ilme
authors.initials: I
authors.lastname: Schlichting
authors.forename: Jochen
authors.initials: J
authors.lastname: Reinstein
mesh_terms: D019943:Amino Acid Substitution; D001426:Bacterial Proteins / Q000737:chemistry / Q000378:metabolism; D018360:Crystallography, X-Ray; D004578:Electron Spin Resonance Spectroscopy; D050956:HSP40 Heat-Shock Proteins / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D008715:Methionine / Q000235:genetics; D008958:Models, Molecular; D018832:Molecular Chaperones / Q000737:chemistry; D011487:Protein Conformation; D017510:Protein Folding; D017434:Protein Structure, Tertiary; D016963:Thermus thermophilus / Q000737:chemistry*
chemical_list: D001426:Bacterial Proteins; D050956:HSP40 Heat-Shock Proteins; D018832:Molecular Chaperones; D008715:Methionine
keywords: cross-linking; crystal dehydration; electron paramagnetic resonance; hybrid structure determination; molecular chaperones; radiation-damage-induced phasing with anomalous scattering; single-wavelength anomalous diffraction
references.citation: Structure. 2000 Aug 15;8(8):799-807
references.pmid: 10997899
references.citation: Acta Crystallogr D Biol Crystallogr. 1998 Sep 1;54(Pt 5):905-21
references.pmid: 9757107
references.citation: FEBS Lett. 1997 Aug 4;412(3):633-6
references.pmid: 9276481
references.citation: J Mol Biol. 1996 Jul 12;260(2):236-50
references.pmid: 8764403
references.citation: J Magn Reson. 2000 Feb;142(2):331-40
references.pmid: 10648151
references.citation: Phys Chem Chem Phys. 2007 Apr 28;9(16):1895-910
references.pmid: 17431518
references.citation: J Biol Chem. 1996 Jul 19;271(29):17343-8
references.pmid: 8663379
references.citation: Acta Crystallogr D Biol Crystallogr. 2004 Nov;60(Pt 11):1958-63
references.pmid: 15502302
references.citation: FEBS Lett. 2005 Oct 24;579(25):5713-7
references.pmid: 16225874
references.citation: Biochemistry. 1999 Mar 30;38(13):4165-76
references.pmid: 10194333
references.citation: J Mol Biol. 1993 Jan 20;229(2):472-93
references.pmid: 8429558
references.citation: Acta Crystallogr D Biol Crystallogr. 2005 Apr;61(Pt 4):458-64
references.pmid: 15805601
references.citation: J Biol Chem. 1994 Oct 28;269(43):27074-9
references.pmid: 7929450
references.citation: J Mol Biol. 2010 May 28;399(1):154-67
references.pmid: 20381501
references.citation: Acta Crystallogr D Biol Crystallogr. 2004 Jun;60(Pt 6):1085-93
references.pmid: 15159568
references.citation: Trends Biochem Sci. 1992 Apr;17(4):129
references.pmid: 1585456
references.citation: J Struct Biol. 1999 Apr-May;125(2-3):156-65
references.pmid: 10222271
references.citation: FEBS Lett. 2009 Feb 4;583(3):573-8
references.pmid: 19162025
references.citation: J Biol Chem. 1995 Feb 3;270(5):2139-44
references.pmid: 7836443
references.citation: Structure. 2003 Feb;11(2):217-24
references.pmid: 12575941
references.citation: Methods Enzymol. 1997;276:523-30
references.pmid: 9048379
references.citation: PLoS One. 2011;6(7):e22337
references.pmid: 21799829
references.citation: Biochemistry. 1999 Aug 10;38(32):10567-77
references.pmid: 10441154
references.citation: J Biol Chem. 1994 Feb 18;269(7):5446-51
references.pmid: 8106526
references.citation: J Biol Chem. 2002 Dec 6;277(49):47160-6
references.pmid: 12351638
references.citation: Acta Crystallogr D Biol Crystallogr. 2011 Apr;67(Pt 4):355-67
references.pmid: 21460454
references.citation: J Mol Biol. 2004 Jun 18;339(5):1179-89
references.pmid: 15178257
references.citation: Angew Chem Int Ed Engl. 1998 Nov 2;37(20):2833-2837
references.pmid: 29711097
references.citation: Cell Mol Life Sci. 2006 Nov;63(22):2560-70
references.pmid: 16952052
references.citation: Acta Crystallogr D Biol Crystallogr. 2005 Sep;61(Pt 9):1289-98
references.pmid: 16131763
references.citation: J Synchrotron Radiat. 2004 Jan 1;11(Pt 1):49-52
references.pmid: 14646132
references.citation: J Biol Chem. 1998 Mar 20;273(12):6643-9
references.pmid: 9506960
references.citation: Cell Mol Life Sci. 2002 Oct;59(10):1598-606
references.pmid: 12475169
references.citation: Acta Crystallogr D Biol Crystallogr. 2002 Oct;58(Pt 10 Pt 2):1772-9
references.pmid: 12351820
references.citation: J Biol Chem. 2005 Nov 18;280(46):38767-75
references.pmid: 16157581
references.citation: Methods Enzymol. 1997;276:307-26
references.pmid: 27754618
references.citation: J Biol Chem. 2003 May 23;278(21):19038-43
references.pmid: 12654915
references.citation: Genetics. 2005 Apr;169(4):1873-82
references.pmid: 15687271
references.citation: Annu Rev Biochem. 1993;62:349-84
references.pmid: 8102520
references.citation: Proc Natl Acad Sci U S A. 2011 Nov 22;108(47):18966-71
references.pmid: 22065753
references.citation: J Mol Biol. 1999 Apr 2;287(3):511-25
references.pmid: 10092456
references.citation: Mol Cell Biol. 1999 Nov;19(11):7751-8
references.pmid: 10523664
references.citation: Acta Crystallogr D Biol Crystallogr. 2010 Apr;66(Pt 4):486-501
references.pmid: 20383002
references.citation: J Biol Chem. 2004 Apr 16;279(16):15723-7
references.pmid: 14729677
references.citation: Proc Natl Acad Sci U S A. 2012 Mar 27;109(13):E734; author reply E735
references.pmid: 22403069
references.citation: Protein Sci. 1999 Jan;8(1):203-14
references.pmid: 10210198
references.citation: Acta Crystallogr D Biol Crystallogr. 2004 Dec;60(Pt 12 Pt 1):2126-32
references.pmid: 15572765
references.citation: J Mol Biol. 2001 Feb 2;305(5):1173-83
references.pmid: 11162122
references.citation: J Magn Reson. 2006 Jan;178(1):42-55
references.pmid: 16188474
references.citation: J Biol Chem. 1996 May 10;271(19):11236-46
references.pmid: 8626673
references.citation: Methods Mol Biol. 2007;364:215-30
references.pmid: 17172768
references.citation: Methods Enzymol. 2003;374:22-37
references.pmid: 14696367
references.citation: J Mol Graph Model. 2002 Dec;21(3):181-3
references.pmid: 12463636
references.citation: Proc Natl Acad Sci U S A. 1994 Nov 22;91(24):11343-7
references.pmid: 7972061
references.citation: Proc Natl Acad Sci U S A. 1993 Nov 1;90(21):10216-20
references.pmid: 8234279
references.citation: Mol Biol Cell. 1993 Jun;4(6):555-63
references.pmid: 8374166
references.citation: Biochemistry. 2010 Oct 5;49(39):8577-84
references.pmid: 20809635
references.citation: Proc Natl Acad Sci U S A. 1999 May 11;96(10):5452-7
references.pmid: 10318904
references.citation: EMBO Rep. 2004 Jun;5(6):567-71
references.pmid: 15170475
references.citation: Trends Biochem Sci. 1994 Apr;19(4):176-81
references.pmid: 8016869
references.citation: Acta Crystallogr A. 2008 Jan;64(Pt 1):112-22
references.pmid: 18156677
references.citation: Trends Cell Biol. 1995 May;5(5):207-12
references.pmid: 14731451
references.citation: Mol Cell. 2007 Nov 9;28(3):422-33
references.pmid: 17996706
references.citation: BMC Struct Biol. 2008 Jan 22;8:3
references.pmid: 18211704
references.citation: Biochem J. 2006 Sep 15;398(3):353-60
references.pmid: 16737444
references.citation: Phys Chem Chem Phys. 2011 Feb 14;13(6):2356-66
references.pmid: 21116569
references.citation: Proteins. 1989;6(3):294-305
references.pmid: 2560193