Structure analysis of membrane-reconstituted subunit c-ring of E. coli H+-ATP synthas

		BioNMR > Educational resources > Journal club > Solid-state high-res. NMR
Structure analysis of membrane-reconstituted subunit c-ring of E. coli H+-ATP synthas

Webservers

NMR processing:

MDD

NMR assignment:

Backbone:

Side-chains:

NOEs:

Structure from NMR restraints:

Ab initio:

Fragment-based:

Rosetta-NMR (Robetta)

Template-based:

GeNMR

I-TASSER

Refinement:

Amber

Structure from chemical shifts:

Fragment-based:

Homology-based:

Torsion angles from chemical shifts:

Preditor

TALOS

Promega- Proline

Secondary structure from chemical shifts:

CSI (via RCI server)

TALOS

MICS caps, β-turns

d2D

PECAN

Flexibility from chemical shifts:

RCI

Interactions from chemical shifts:

HADDOCK

Chemical shifts re-referencing:

NMR model quality:

NOEs, other restraints:

Chemical shifts:

RDCs:

Pseudocontact shifts:

Protein geomtery:

SAVES2 or SAVES4

Quality Control Check

NMR spectrum prediction:

FANDAS

MestReS

V-NMR

Flexibility from structure:

Backbone S2

Methyl S2

B-factor

Molecular dynamics:

Gromacs

Amber

Antechamber

Chemical shifts prediction:

From structure:

Shiftx2

Sparta+

Camshift

CH3shift- Methyl

ArShift- Aromatic

ShiftS

Proshift

PPM

CheShift-2- Cα

From sequence:

Shifty

Camcoil

Poulsen_rc_CS

Disordered proteins:

MAXOCC

Format conversion & validation:

CCPN

From NMR-STAR 3.1

Validate NMR-STAR 3.1

NMR sample preparation:

Protein disorder:

DisMeta

Protein solubility:

Isotope labeling:

Solid-state NMR:

Thread Tools

Search this Thread

Rating:

Display Modes

08-08-2010, 01:57 AM

nmrlearner

Senior Member

Join Date: Jan 2005

Posts: 23,731

Points: 193,617, Level: 100

Level up: 0%, 0 Points needed

Activity: 50.7%

Last Achievements

Award-Showcase

NMR Credits: 0

NMR Points: 193,617

Downloads: 0

Uploads: 0

Structure analysis of membrane-reconstituted subunit c-ring of E. coli H+-ATP synthas

Abstract The subunit c-ring of H+-ATP synthase (Fo c-ring) plays an essential role in the proton translocation across a membrane driven by the electrochemical potential. To understand its structure and function, we have carried out solid-state NMR analysis under magic-angle sample spinning. The uniformly [13C, 15N]-labeled Fo c from E. coli (EFo c) was reconstituted into lipid membranes as oligomers. Its high resolution two- and three-dimensional spectra were obtained, and the 13C and 15N signals were assigned. The obtained chemical shifts suggested that EFo c takes on a hairpin-type helix-loop-helix structure in membranes as in an organic solution. The results on the magnetization transfer between the EFo c and deuterated lipids indicated that Ile55, Ala62, Gly69 and F76 were lined up on the outer surface of the oligomer. This is in good agreement with the cross-linking results previously reported by Fillingame and his colleagues. This agreement reveals that the reconstituted EFo c oligomer takes on a ring structure similar to the intact one in vivo. On the other hand, analysis of the 13C nuclei distance of [3-13C]Ala24 and [4-13C]Asp61 in the Fo c-ring did not agree with the model structures proposed for the EFo c-decamer and dodecamer. Interestingly, the carboxyl group of the essential Asp61 in the membrane-embedded EFo c-ring turned out to be protonated as COOH even at neutral pH. The hydrophobic surface of the EFo c-ring carries relatively short side chains in its central region, which may allow soft and smooth interactions with the hydrocarbon chains of lipids in the liquid-crystalline state.

Content Type Journal Article
DOI 10.1007/s10858-010-9432-x
Authors
- Yasuto Todokoro, Osaka University Institute for Protein Research 3-2 Yamadaoka Suita 565-0871 Japan
- Masatoshi Kobayashi, Osaka University Institute for Protein Research 3-2 Yamadaoka Suita 565-0871 Japan
- Takeshi Sato, Osaka University Institute for Protein Research 3-2 Yamadaoka Suita 565-0871 Japan
- Toru Kawakami, Osaka University Institute for Protein Research 3-2 Yamadaoka Suita 565-0871 Japan
- Ikuko Yumen, Osaka University Institute for Protein Research 3-2 Yamadaoka Suita 565-0871 Japan
- Saburo Aimoto, Osaka University Institute for Protein Research 3-2 Yamadaoka Suita 565-0871 Japan
- Toshimichi Fujiwara, Osaka University Institute for Protein Research 3-2 Yamadaoka Suita 565-0871 Japan
- Hideo Akutsu, Osaka University Institute for Protein Research 3-2 Yamadaoka Suita 565-0871 Japan

- Journal Journal of Biomolecular NMR
- Online ISSN 1573-5001
- Print ISSN 0925-2738

Source: Journal of Biomolecular NMR

Similar Threads
Thread	Thread Starter	Forum	Replies	Last Post
NMR spectroscopy with the stringent substrate rhodanese bound to the single-ring variant SR1 of the E. coli chaperonin GroEL. NMR spectroscopy with the stringent substrate rhodanese bound to the single-ring variant SR1 of the E. coli chaperonin GroEL. NMR spectroscopy with the stringent substrate rhodanese bound to the single-ring variant SR1 of the E. coli chaperonin GroEL. Protein Sci. 2011 Jun 1; Authors: Koculi E, Horst R, Horwich AL, Wüthrich K NMR observation of the uniformly (2) H,(15) N-labeled stringent 33 kDa substrate protein rhodanese in a productive complex with the uniformly (14) N-labeled 400 kDa single-ring version of the E. coli chaperonin GroEL, SR1,...	nmrlearner	Journal club	0	06-03-2011 10:20 AM
Structure analysis of membrane-reconstituted subunit c-ring of E. coli H+-ATP synthase by solid-state NMR. Structure analysis of membrane-reconstituted subunit c-ring of E. coli H+-ATP synthase by solid-state NMR. Structure analysis of membrane-reconstituted subunit c-ring of E. coli H+-ATP synthase by solid-state NMR. J Biomol NMR. 2010 Sep;48(1):1-11 Authors: Todokoro Y, Kobayashi M, Sato T, Kawakami T, Yumen I, Aimoto S, Fujiwara T, Akutsu H The subunit c-ring of H(+)-ATP synthase (F(o) c-ring) plays an essential role in the proton translocation across a membrane driven by the electrochemical potential. To understand its structure and function, we...	nmrlearner	Journal club	0	12-18-2010 12:00 PM
[NMR paper] Subunit A of the E. coli ATP synthase: reconstitution and high resolution NMR with pr Subunit A of the E. coli ATP synthase: reconstitution and high resolution NMR with protein purified in a mixed polarity solvent. Related Articles Subunit A of the E. coli ATP synthase: reconstitution and high resolution NMR with protein purified in a mixed polarity solvent. FEBS Lett. 2004 Jan 2;556(1-3):35-8 Authors: Dmitriev OY, Altendorf K, Fillingame RH Subunit a of the Escherichia coli ATP synthase, a 30 kDa integral membrane protein, was purified to homogeneity by a novel procedure incorporating selective extraction into a monophasic...	nmrlearner	Journal club	0	11-24-2010 09:25 PM
[NMR paper] NMR solution structure determination of membrane proteins reconstituted in detergent NMR solution structure determination of membrane proteins reconstituted in detergent micelles. Related Articles NMR solution structure determination of membrane proteins reconstituted in detergent micelles. FEBS Lett. 2003 Nov 27;555(1):144-50 Authors: Fernández C, Wüthrich K As an alternative to X-ray crystallography, nuclear magnetic resonance (NMR) spectroscopy in solution can be used for three-dimensional structure determination of small membrane proteins, preferably proteins with beta-barrel fold. This paper reviews recent achievements as...	nmrlearner	Journal club	0	11-24-2010 09:16 PM
[NMR paper] Elucidation of the epsilon-theta subunit interface of Escherichia coli DNA polymerase Elucidation of the epsilon-theta subunit interface of Escherichia coli DNA polymerase III by NMR spectroscopy. Related Articles Elucidation of the epsilon-theta subunit interface of Escherichia coli DNA polymerase III by NMR spectroscopy. Biochemistry. 2003 Apr 8;42(13):3635-44 Authors: DeRose EF, Darden T, Harvey S, Gabel S, Perrino FW, Schaaper RM, London RE The DNA polymerase III holoenzyme (HE) is the primary replicative polymerase of Escherichia coli. The epsilon (epsilon) subunit of HE provides the 3'-->5' exonucleolytic proofreading...	nmrlearner	Journal club	0	11-24-2010 09:01 PM
[NMR paper] Side chain NMR assignments in the membrane protein OmpX reconstituted in DHPC micelle Side chain NMR assignments in the membrane protein OmpX reconstituted in DHPC micelles. Related Articles Side chain NMR assignments in the membrane protein OmpX reconstituted in DHPC micelles. J Biomol NMR. 2002 Aug;23(4):289-301 Authors: Hilty C, Fernández C, Wider G, Wüthrich K Sequence-specific assignments have been obtained for side chain methyl resonances of Val, Leu and Ile in the outer membrane protein X (OmpX) from Escherichia coli reconstituted in 60 kDa micelles in aqueous solution. Using previously established techniques, OmpX was...	nmrlearner	Journal club	0	11-24-2010 08:58 PM
[NMR paper] Structural features of the epsilon subunit of the Escherichia coli ATP synthase deter Structural features of the epsilon subunit of the Escherichia coli ATP synthase determined by NMR spectroscopy. Related Articles Structural features of the epsilon subunit of the Escherichia coli ATP synthase determined by NMR spectroscopy. Nat Struct Biol. 1995 Nov;2(11):961-7 Authors: Wilkens S, Dahlquist FW, McIntosh LP, Donaldson LW, Capaldi RA The tertiary fold of the epsilon subunit of the Escherichia coli F1F0 ATPsynthase (ECF1F0) has been determined by two- and three-dimensional heteronuclear (13C, 15N) NMR spectroscopy. The epsilon...	nmrlearner	Journal club	0	08-22-2010 03:50 AM
[NMR paper] Proton NMR of Escherichia coli sulfite reductase: the unligated hemeprotein subunit. Proton NMR of Escherichia coli sulfite reductase: the unligated hemeprotein subunit. Related Articles Proton NMR of Escherichia coli sulfite reductase: the unligated hemeprotein subunit. Biochemistry. 1993 Mar 23;32(11):2853-67 Authors: Kaufman J, Spicer LD, Siegel LM The isolated hemeprotein subunit of sulfite reductase (SiR-HP) from Escherichia coli consists of a high spin ferric isobacteriochlorin (siroheme) coupled to a diamagnetic 2+ cluster. When supplied with an artificial electron donor, such as methyl viologen cation radical, SiR-HP...	nmrlearner	Journal club	0	08-21-2010 11:53 PM

« Previous Thread | Next Thread »

Posting Rules
You may not post new threads You may not post replies You may not post attachments You may not edit your posts BB code is On Smilies are On [IMG] code is On HTML code is On Trackbacks are Off Pingbacks are Off Refbacks are Off