NMR paper Two-dimensional 1H, 15N NMR investigation of uniformly 15N-labeled lac repressor head

		BioNMR > Educational resources > Journal club
[NMR paper] Two-dimensional 1H, 15N NMR investigation of uniformly 15N-labeled lac repressor head

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

08-21-2010, 11:41 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

Two-dimensional 1H, 15N NMR investigation of uniformly 15N-labeled lac repressor head

Two-dimensional 1H, 15N NMR investigation of uniformly 15N-labeled lac repressor headpiece.

Related Articles Two-dimensional 1H, 15N NMR investigation of uniformly 15N-labeled lac repressor headpiece.

J Biomol Struct Dyn. 1992 Apr;9(5):921-33

Authors: Lancelot G, Gervais A, Maurizot JC

15N uniformly labeled lac repressor and lac repressor headpiece were prepared. 15N NMR spectra of lac repressor were shown resolution inadequate for detailed study while the data showed that the 15N labeled N-terminal part of the protein is quite suitable for this type of study allowing future investigation of the specific interaction of the lac repressor headpiece with the lac operator. We report here the total assignment of proton 1H and nitrogen 15NH backbone resonances of this headpiece in the free state. Assignments of the 15N resonances of the protein were obtained in a sequential manner using heteronuclear multiple quantum coherence (HMQC), relayed HMQC nuclear Overhauser and relayed HMQC-HOHAHA spectroscopy. More than 80 per cent of residues were assigned by their 15NH(i)-N1H(i + 1) and 15NH(i)-N1H(i - 1) connectivities. Values of the 3JNH alpha splitting for 39 of the 51 residues of the headpiece were extracted from HMQC and HMQC-J. The observed 15NH(i)-C beta H cross peaks and the 3JNH alpha coupling constants values are in agreement with the three alpha-helices previously described [Zuiderweg, E.R.P., Scheek, R.M., Boelens, R., van Gunsteren, W.F. and Kaptein, R., Biochimie 67, 707 (1985)]. The 3JNH alpha coupling constants can be now used for a more confident determination of the lac repressor headpiece. From these values it is shown that the geometry of the ends of the second and third alpha-helices exhibit deviation from the canonical alpha-helix structure. On the basis of NOEs and 3JNH alpha values, the geometry of the turn of the helix-turn-helix motif is discussed.

PMID: 1524707 [PubMed - indexed for MEDLINE]

Source: PubMed

Did you find this post helpful?

Similar Threads
Thread	Thread Starter	Forum	Replies	Last Post
[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] Towards high-resolution solid-state NMR on large uniformly 15N- and [13C,15N]-labeled Towards high-resolution solid-state NMR on large uniformly 15N- and -labeled membrane proteins in oriented lipid bilayers. Related Articles Towards high-resolution solid-state NMR on large uniformly 15N- and -labeled membrane proteins in oriented lipid bilayers. J Biomol NMR. 2002 Mar;22(3):225-47 Authors: Vosegaard T, Nielsen NC Based on exact numerical simulations, taking into account isotropic and conformation-dependent anisotropic nuclear spin interactions, we systematically analyse the prospects for high-resolution solid-state NMR on...	nmrlearner	Journal club	0	11-24-2010 08:49 PM
[NMR paper] Complete resolution of the solid-state NMR spectrum of a uniformly 15N-labeled membra Complete resolution of the solid-state NMR spectrum of a uniformly 15N-labeled membrane protein in phospholipid bilayers. http://www.ncbi.nlm.nih.gov/corehtml/query/egifs/http:--highwire.stanford.edu-icons-externalservices-pubmed-custom-pnas_full_free.gif http://www.ncbi.nlm.nih.gov/corehtml/query/egifs/http:--www.pubmedcentral.nih.gov-corehtml-pmc-pmcgifs-pubmed-pmc.gif Related Articles Complete resolution of the solid-state NMR spectrum of a uniformly 15N-labeled membrane protein in phospholipid bilayers. Proc Natl Acad Sci U S A. 1997 Aug 5;94(16):8551-6 ...	nmrlearner	Journal club	0	08-22-2010 05:08 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
[NMR paper] Expression of human chorionic gonadotropin uniformly labeled with NMR isotopes in Chi Expression of human chorionic gonadotropin uniformly labeled with NMR isotopes in Chinese hamster ovary cells: an advance toward rapid determination of glycoprotein structures. Related Articles Expression of human chorionic gonadotropin uniformly labeled with NMR isotopes in Chinese hamster ovary cells: an advance toward rapid determination of glycoprotein structures. J Biomol NMR. 1996 Jun;7(4):295-304 Authors: Lustbader JW, Birken S, Pollak S, Pound A, Chait BT, Mirza UA, Ramnarain S, Canfield RE, Brown JM Most secreted eukaryotic proteins...	nmrlearner	Journal club	0	08-22-2010 02:27 PM
[NMR paper] 15N NMR assignments and chemical shift analysis of uniformly labeled 15N calbindin D9 15N NMR assignments and chemical shift analysis of uniformly labeled 15N calbindin D9k in the apo, (Cd2+)1 and (Ca2+)2 states. Related Articles 15N NMR assignments and chemical shift analysis of uniformly labeled 15N calbindin D9k in the apo, (Cd2+)1 and (Ca2+)2 states. FEBS Lett. 1992 Jun 1;303(2-3):136-40 Authors: Skelton NJ, Akke M, Kördel J, Thulin E, Forsén S, Chazin WJ 15N has been uniformly incorporated into the EF-hand Ca(2+)-binding protein calbindin D9k so that heteronuclear experiments can be used to further characterize the...	nmrlearner	Journal club	0	08-21-2010 11:41 PM
1H-13C Separated Local Field Spectroscopy of Uniformly 13C Labeled Peptides and Prote 1H-13C Separated Local Field Spectroscopy of Uniformly 13C Labeled Peptides and Proteins Publication year: 2010 Source: Journal of Magnetic Resonance, In Press, Accepted Manuscript, Available online 1 July 2010</br> Eugene C., Lin , Chin H., Wu , Yuan, Yang , Christopher V., Grant , Stanley J., Opella</br> By incorporating homonuclear decoupling on both the 1H and 13C channels it is feasible to obtain high-resolution two-dimensional separated local field spectra of peptides and proteins that are 100% labeled with 13C. Dual-PISEMO (Polarization Inversion Spin Exchange Modulated...	nmrlearner	Journal club	0	08-16-2010 03:50 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