NMR paper Cost-effective method for the preparation of uniformly labeled myristoylated proteins for NMR measurements.

		BioNMR > Educational resources > Journal club
[NMR paper] Cost-effective method for the preparation of uniformly labeled myristoylated proteins for NMR measurements.

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

03-26-2014, 12:44 PM

nmrlearner

Senior Member

Join Date: Jan 2005

Posts: 23,777

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

Cost-effective method for the preparation of uniformly labeled myristoylated proteins for NMR measurements.

Cost-effective method for the preparation of uniformly labeled myristoylated proteins for NMR measurements.

Related Articles Cost-effective method for the preparation of uniformly labeled myristoylated proteins for NMR measurements.

Protein Expr Purif. 2014 Mar 21;

Authors: Kroupa T, Prchal J, Doležal M, Ruml T, Hrabal R

Abstract
Nuclear magnetic resonance (NMR) is a powerful technique for solving protein structures orstudying their interactions. However, it requires molecules labeled with NMR sensitive isotopes like carbon(13)C and nitrogen(15)N. The recombinant expression of labeled proteins is simple to perform but requires quite expensive chemicals. When there is a need for special labeled chemicals, like uniformly (13)C-labeled myristic acid, the price significantly rises. Here we describe a cost-effective method for the recombinant expression ofuniformly labeled myristoylated proteins in Escherichia coli demonstrated on the production of Mason-Pfizer monkeyvirus matrix protein.We used the ability of E. coli to naturally synthetize myristic acid. Whengrown in isotopically labeled medium the myristic acid will be labelled as well. Bacteria wereco-transfected with plasmid carrying gene for yeast N-myristoyltransferase which ensures myristoylation of expressed protein. This process provided1.8 mgof the myristoylated, doubly labeled ((13)C/(15)N)M-PMV matrix proteinfrom 1 l of (15)N/(13)C labeled M9 medium. The price represents approximately 50% cost reduction of conventional method using commercially available [U-(13)C]myristic acid.

PMID: 24662511 [PubMed - as supplied by publisher]

More...

Did you find this post helpful?

Similar Threads
Thread	Thread Starter	Forum	Replies	Last Post
Cost-effective method for the preparation of uniformly labeled myristoylated proteins for NMR measurements Cost-effective method for the preparation of uniformly labeled myristoylated proteins for NMR measurements Publication date: Available online 21 March 2014 Source:Protein Expression and Purification</br> Author(s): Tomáš Kroupa , Jan Prchal , Michal Doležal , Tomáš Ruml , Richard Hrabal</br> Nuclear magnetic resonance (NMR) is a powerful technique for solving protein structures orstudying their interactions. However, it requires molecules labeled with NMR sensitive isotopes like carbon13C and nitrogen15N. The recombinant expression of labeled proteins is simple...	nmrlearner	Journal club	0	03-22-2014 01:28 AM
[NMR paper] A Cost-Effective Protocol for the Parallel Production of Libraries of (13)CH 3-Specifically Labeled Mutants for NMR Studies of High Molecular Weight Proteins. A Cost-Effective Protocol for the Parallel Production of Libraries of (13)CH 3-Specifically Labeled Mutants for NMR Studies of High Molecular Weight Proteins. Related Articles A Cost-Effective Protocol for the Parallel Production of Libraries of (13)CH 3-Specifically Labeled Mutants for NMR Studies of High Molecular Weight Proteins. Methods Mol Biol. 2014;1091:229-44 Authors: Crublet E, Kerfah R, Mas G, Noirclerc-Savoye M, Lantez V, Vernet T, Boisbouvier J Abstract There is increasing interest in applying NMR spectroscopy to the study of...	nmrlearner	Journal club	0	11-11-2013 01:30 AM
A cost-effective look at proteins - with a little help from robotics - Cordis News http://www.bionmr.com//t3.gstatic.com/images?q=tbn:ANd9GcTypMhbcsoTBmkGzm1ZmzjTHj-b-AsNVL4y8L03Y0oBfj2cCZUDv2gZmGEzmim-0VUKjKStcwU Cordis News <img alt="" height="1" width="1" /> A cost-effective look at proteins - with a little help from robotics Cordis News Until now, a number of methods have been used to study static protein structures, including X-ray crystallography and nuclear magnetic resonance. However, these methods are of no use for proteins that are on the move; analytical methods, computer ... and more » A cost-effective look at proteins - with a little help from...	nmrlearner	Online News	0	09-14-2013 01:46 AM
[NMR paper] Preparation of uniformly isotope labeled KcsA for solid state NMR: Expression, purification, reconstitution into liposomes and functional assay. Preparation of uniformly isotope labeled KcsA for solid state NMR: Expression, purification, reconstitution into liposomes and functional assay. Preparation of uniformly isotope labeled KcsA for solid state NMR: Expression, purification, reconstitution into liposomes and functional assay. Protein Expr Purif. 2013 Aug 1; Authors: Bhate MP, Wylie BJ, Thompson A, Tian L, Nimigean C, McDermott AE Abstract We report the expression, purification, liposome reconstitution and functional validation of uniformly (13)C and (15)N isotope labeled...	nmrlearner	Journal club	0	08-07-2013 01:24 PM
[NMR paper] Site-specific 13C chemical shift anisotropy measurements in a uniformly 15N,13C-labeled microcrystalline protein by 3D magic-angle spinning NMR spectroscopy. Site-specific 13C chemical shift anisotropy measurements in a uniformly 15N,13C-labeled microcrystalline protein by 3D magic-angle spinning NMR spectroscopy. Related Articles Site-specific 13C chemical shift anisotropy measurements in a uniformly 15N,13C-labeled microcrystalline protein by 3D magic-angle spinning NMR spectroscopy. J Am Chem Soc. 2005 Aug 31;127(34):11946-7 Authors: Wylie BJ, Franks WT, Graesser DT, Rienstra CM In this Communication, we introduce a 3D magic-angle spinning recoupling experiment that correlates chemical shift...	nmrlearner	Journal club	0	12-01-2010 06:56 PM
[NMR paper] Preparation of uniformly labeled NMR samples in Escherichia coli under the tight cont Preparation of uniformly labeled NMR samples in Escherichia coli under the tight control of the araBAD promoter: expression of an archaeal homolog of the RNase P Rpp29 protein. Related Articles Preparation of uniformly labeled NMR samples in Escherichia coli under the tight control of the araBAD promoter: expression of an archaeal homolog of the RNase P Rpp29 protein. Protein Expr Purif. 2003 Apr;28(2):246-51 Authors: Boomershine WP, Raj ML, Gopalan V, Foster MP We report the first use of the tightly regulated araBAD promoter for generating...	nmrlearner	Journal club	0	11-24-2010 09:01 PM
[NMR paper] Preparation and characterization of a uniformly 2 H/ 15 N-labeled RNA oligonucleotide Preparation and characterization of a uniformly 2 H/ 15 N-labeled RNA oligonucleotide for NMR studies. http://www.ncbi.nlm.nih.gov/corehtml/query/egifs/http:--highwire.stanford.edu-icons-externalservices-pubmed-custom-oxfordjournals_final_free.gif http://www.ncbi.nlm.nih.gov/corehtml/query/egifs/http:--www.pubmedcentral.nih.gov-corehtml-pmc-pmcgifs-pubmed-pmc.gif Related Articles Preparation and characterization of a uniformly 2 H/ 15 N-labeled RNA oligonucleotide for NMR studies. Nucleic Acids Res. 1997 Apr 1;25(7):1390-6 Authors: Nikonowicz EP,...	nmrlearner	Journal club	0	08-22-2010 03:31 PM
[NMR paper] Preparation and characterization of a uniformly 2 H/ 15 N-labeled RNA oligonucleotide Preparation and characterization of a uniformly 2 H/ 15 N-labeled RNA oligonucleotide for NMR studies. http://www.ncbi.nlm.nih.gov/corehtml/query/egifs/http:--highwire.stanford.edu-icons-externalservices-pubmed-custom-oxfordjournals_final_free.gif http://www.ncbi.nlm.nih.gov/corehtml/query/egifs/http:--www.pubmedcentral.nih.gov-corehtml-pmc-pmcgifs-pubmed-pmc.gif Related Articles Preparation and characterization of a uniformly 2 H/ 15 N-labeled RNA oligonucleotide for NMR studies. Nucleic Acids Res. 1997 Apr 1;25(7):1390-6 Authors: Nikonowicz EP,...	nmrlearner	Journal club	0	08-22-2010 03:03 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