NMR paper Membrane protein preparation for TROSY NMR screening.

		BioNMR > Educational resources > Journal club
[NMR paper] Membrane protein preparation for TROSY NMR screening.

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

Rate Thread

Display Modes

11-24-2010, 11:14 PM

nmrlearner

Senior Member

Join Date: Jan 2005

Posts: 23,697

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

Membrane protein preparation for TROSY NMR screening.

Membrane protein preparation for TROSY NMR screening.

Related Articles Membrane protein preparation for TROSY NMR screening.

Methods Enzymol. 2005;394:321-34

Authors: Tian C, Karra MD, Ellis CD, Jacob J, Oxenoid K, Sönnichsen F, Sanders CR

The first steps toward undertaking an NMR structural study of a new protein is very often to purify the protein and then to acquire an HSQC or TROSY NMR spectrum, the quality of which is used to assess the feasibility of an NMR-based structural determination. Relatively few integral membrane proteins (IMPs) have been subjected even to this very preliminary stage of NMR analysis. Here, NMR feasibility testing methods are outlined that are tailored for hexahistidine-tagged IMPs that have been expressed in Escherichia coli. Generally applicable protocols are presented for expression testing, purification, and NMR sample preparation. A 2D TROSY pulse sequence that has been optimized for use with IMPs is also presented.

PMID: 15808226 [PubMed - indexed for MEDLINE]

Source: PubMed

Did you find this post helpful?

Similar Threads
Thread	Thread Starter	Forum	Replies	Last Post
Preparation of protein samples for NMR structure, function, and small-molecule screening studies. Preparation of protein samples for NMR structure, function, and small-molecule screening studies. Preparation of protein samples for NMR structure, function, and small-molecule screening studies. Methods Enzymol. 2011;493:21-60 Authors: Acton TB, Xiao R, Anderson S, Aramini J, Buchwald WA, Ciccosanti C, Conover K, Everett J, Hamilton K, Huang YJ, Janjua H, Kornhaber G, Lau J, Lee DY, Liu G, Maglaqui M, Ma L, Mao L, Patel D, Rossi P, Sahdev S, Shastry R, Swapna GV, Tang Y, Tong S, Wang D, Wang H, Zhao L, Montelione GT In this chapter, we...	nmrlearner	Journal club	0	03-05-2011 01:02 PM
Solid-State NMR on a Large Multidomain Integral Membrane Protein: The Outer Membrane Protein Assembly Factor BamA. Solid-State NMR on a Large Multidomain Integral Membrane Protein: The Outer Membrane Protein Assembly Factor BamA. Solid-State NMR on a Large Multidomain Integral Membrane Protein: The Outer Membrane Protein Assembly Factor BamA. J Am Chem Soc. 2011 Mar 1; Authors: Renault M, Bos MP, Tommassen J, Baldus M Multidomain proteins constitute a large part of prokaryotic and eukaryotic proteomes and play fundamental roles in various physiological processes. However, their structural characterization is challenging because of their large size and...	nmrlearner	Journal club	0	03-03-2011 12:34 PM
Solid-State NMR on a Large Multidomain Integral Membrane Protein: The Outer Membrane Protein Assembly Factor BamA Solid-State NMR on a Large Multidomain Integral Membrane Protein: The Outer Membrane Protein Assembly Factor BamA Marie Renault, Martine P. Bos, Jan Tommassen and Marc Baldus http://pubs.acs.org/appl/literatum/publisher/achs/journals/content/jacsat/0/jacsat.ahead-of-print/ja109469c/aop/images/medium/ja-2010-09469c_0004.gif Journal of the American Chemical Society DOI: 10.1021/ja109469c http://feeds.feedburner.com/~ff/acs/jacsat?d=yIl2AUoC8zA http://feeds.feedburner.com/~r/acs/jacsat/~4/9XN1qiW-S-I	nmrlearner	Journal club	0	03-02-2011 02:01 AM
[NMR paper] NMR structure of Mistic, a membrane-integrating protein for membrane protein expressi NMR structure of Mistic, a membrane-integrating protein for membrane protein expression. Related Articles NMR structure of Mistic, a membrane-integrating protein for membrane protein expression. Science. 2005 Feb 25;307(5713):1317-21 Authors: Roosild TP, Greenwald J, Vega M, Castronovo S, Riek R, Choe S Although structure determination of soluble proteins has become routine, our understanding of membrane proteins has been limited by experimental bottlenecks in obtaining both sufficient yields of protein and ordered crystals. Mistic is an...	nmrlearner	Journal club	0	11-24-2010 11:14 PM
[NMR paper] 3D TROSY-HNCA(coded)CB and TROSY-HNCA(coded)CO experiments: triple resonance NMR expe 3D TROSY-HNCA(coded)CB and TROSY-HNCA(coded)CO experiments: triple resonance NMR experiments with two sequential connectivity pathways and high sensitivity. Related Articles 3D TROSY-HNCA(coded)CB and TROSY-HNCA(coded)CO experiments: triple resonance NMR experiments with two sequential connectivity pathways and high sensitivity. J Biomol NMR. 2004 Mar;28(3):289-94 Authors: Ritter C, Lührs T, Kwiatkowski W, Riek R The concept of chemical shift-coding monitors chemical shifts in multi-dimensional NMR experiments without additional polarization...	nmrlearner	Journal club	0	11-24-2010 09:25 PM
[NMR paper] Preparation of protein nanocrystals and their characterization by solid state NMR. Preparation of protein nanocrystals and their characterization by solid state NMR. Related Articles Preparation of protein nanocrystals and their characterization by solid state NMR. J Magn Reson. 2003 Nov;165(1):162-74 Authors: Martin RW, Zilm KW Preparation of proteins in their crystalline state has been found to be important in producing stable therapeutic protein formulations, cross-linked enzyme crystals for application in industrial processes, generating novel porous media for separations, and of course in structure elucidation. Of these...	nmrlearner	Journal club	0	11-24-2010 09:16 PM
[NMR paper] Protein dynamics measurements by TROSY-based NMR experiments. Protein dynamics measurements by TROSY-based NMR experiments. Related Articles Protein dynamics measurements by TROSY-based NMR experiments. J Magn Reson. 2000 Apr;143(2):423-6 Authors: Zhu G, Xia Y, Nicholson LK, Sze KH The described TROSY-based experiments for investigating backbone dynamics of proteins make it possible to elucidate internal motions in large proteins via measurements of T(1), T(2), and NOE of backbone (15)N nuclei. In our proposed sequences, the INEPT sequence is eliminated and the PEP sequence is replaced by the ST2-PT...	nmrlearner	Journal club	0	11-18-2010 09:15 PM
Simultaneous Structure and Dynamics of a Membrane Protein using REDCRAFT: Membrane-bo Simultaneous Structure and Dynamics of a Membrane Protein using REDCRAFT: Membrane-bound form of Pf1 Coat Protein Publication year: 2010 Source: Journal of Magnetic Resonance, In Press, Accepted Manuscript, Available online 30 July 2010</br> Paul, Shealy , Mikhail, Simin , Sang Ho, Park , Stanley J., Opella , Homayoun, Valafar</br> A strategy for simultaneous study of the structure and internal dynamics of a membrane protein is described using the REDCRAFT algorithm. The membrane-bound form of the Pf1 major coat protein (mbPf1) was used as an example. First, synthetic data is...	nmrlearner	Journal club	0	08-15-2010 12:32 AM

« 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