Interfacial enzyme kinetics of a membrane bound kinase analyzed by real-time MAS-NMR.

		BioNMR > Educational resources > Journal club
Interfacial enzyme kinetics of a membrane bound kinase analyzed by real-time MAS-NMR.

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

03-23-2011, 05:41 PM

nmrlearner

Senior Member

Join Date: Jan 2005

Posts: 23,732

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

Interfacial enzyme kinetics of a membrane bound kinase analyzed by real-time MAS-NMR.

Interfacial enzyme kinetics of a membrane bound kinase analyzed by real-time MAS-NMR.

Interfacial enzyme kinetics of a membrane bound kinase analyzed by real-time MAS-NMR.

Nat Chem Biol. 2011 Mar 20;

Authors: Ullrich SJ, Hellmich UA, Ullrich S, Glaubitz C

The simultaneous observation of interdependent reactions within different phases as catalyzed by membrane-bound enzymes is still a challenging task. One such enzyme, the Escherichia coli integral membrane protein diacylglycerol kinase (DGK), is a key player in lipid regulation. It catalyzes the generation of phosphatidic acid within the membrane through the transfer of the ?-phosphate from soluble MgATP to membrane-bound diacylglycerol. We demonstrate that time-resolved (31)P magic angle spinning NMR offers a unique opportunity to simultaneously and directly detect both ATP hydrolysis and diacylglycerol phosphorylation. This experiment demonstrates that solid-state NMR provides a general approach for the kinetic analysis of coupled reactions at the membrane interface regardless of their compartmentalization. The enzymatic activity of DGK was probed with different lipid substrates as well as ATP analogs. Our data yield conclusions about intersubunit cooperativity, reaction stoichiometries and phosphoryl transfer mechanism and are discussed in the context of known structural data.

PMID: 21423170 [PubMed - as supplied by publisher]

Source: PubMed

Did you find this post helpful?

Similar Threads
Thread	Thread Starter	Forum	Replies	Last Post
Transient Enzyme–Substrate Recognition Monitored by Real-Time NMR Transient Enzyme–Substrate Recognition Monitored by Real-Time NMR Caroline Haupt, Rica Patzschke, Ulrich Weininger, Stefan Gro?ger, Michael Kovermann and Jochen Balbach http://pubs.acs.org/appl/literatum/publisher/achs/journals/content/jacsat/0/jacsat.ahead-of-print/ja2010048/aop/images/medium/ja-2011-010048_0002.gif Journal of the American Chemical Society DOI: 10.1021/ja2010048 http://feeds.feedburner.com/~ff/acs/jacsat?d=yIl2AUoC8zA http://feeds.feedburner.com/~r/acs/jacsat/~4/nknzYbs0FNE	nmrlearner	Journal club	0	06-30-2011 05:01 AM
Transient enzyme-substrate recognition monitored by real-time NMR. Transient enzyme-substrate recognition monitored by real-time NMR. Transient enzyme-substrate recognition monitored by real-time NMR. J Am Chem Soc. 2011 Jun 10; Authors: Haupt C, Patzschke R, Weininger U, Gröger S, Kovermann M, Balbach J Slow protein folding processes during which kinetic folding intermediates occur for an extended time can lead to aggregation and dysfunction in living cells. Therefore protein folding helpers have evolved, which prevent proteins from aggregation and/ or speed up folding processes. In this study we present the...	nmrlearner	Journal club	0	06-15-2011 01:15 PM
[NMR paper] How to prepare membrane proteins for solid-state NMR: A case study on the alpha-helical integral membrane protein diacylglycerol kinase from E. coli. How to prepare membrane proteins for solid-state NMR: A case study on the alpha-helical integral membrane protein diacylglycerol kinase from E. coli. Related Articles How to prepare membrane proteins for solid-state NMR: A case study on the alpha-helical integral membrane protein diacylglycerol kinase from E. coli. Chembiochem. 2005 Sep;6(9):1693-700 Authors: Lorch M, Faham S, Kaiser C, Weber I, Mason AJ, Bowie JU, Glaubitz C Several studies have demonstrated that it is viable to use microcrystalline preparations of water-soluble proteins as...	nmrlearner	Journal club	0	12-01-2010 06:56 PM
[NMR paper] Protein folding studied by real-time NMR spectroscopy. Protein folding studied by real-time NMR spectroscopy. Related Articles Protein folding studied by real-time NMR spectroscopy. Methods. 2004 Sep;34(1):65-74 Authors: Zeeb M, Balbach J Real-time NMR spectroscopy developed to a generally applicable method to follow protein folding reactions. It combines the access to high resolution data with kinetic experiments allowing very detailed insights into the development of the protein structure during different steps of folding. The present review concentrates mainly on the progress of real-time NMR...	nmrlearner	Journal club	0	11-24-2010 10:01 PM
[NMR paper] Folding kinetics of the SH3 domain of PI3 kinase by real-time NMR combined with optic Folding kinetics of the SH3 domain of PI3 kinase by real-time NMR combined with optical spectroscopy. Related Articles Folding kinetics of the SH3 domain of PI3 kinase by real-time NMR combined with optical spectroscopy. J Mol Biol. 1998 Feb 27;276(3):657-67 Authors: Guijarro JI, Morton CJ, Plaxco KW, Campbell ID, Dobson CM The refolding kinetics of the chemically denatured SH3 domain of phosphatidylinositol 3'-kinase (PI3-SH3) have been monitored by real-time one-dimensional 1H NMR coupled with a variety of other biophysical techniques. These...	nmrlearner	Journal club	0	11-17-2010 11:06 PM
[NMR paper] Following protein folding in real time using NMR spectroscopy. Following protein folding in real time using NMR spectroscopy. Related Articles Following protein folding in real time using NMR spectroscopy. Nat Struct Biol. 1995 Oct;2(10):865-70 Authors: Balbach J, Forge V, van Nuland NA, Winder SL, Hore PJ, Dobson CM The refolding of apo bovine alpha-lactalbumin has been monitored in real time by NMR spectroscopy following rapid in situ dilution of a chemically denatured state. By examining individual resonances in the time-resolved NMR spectra, the native state has been shown to emerge in a cooperative...	nmrlearner	Journal club	0	08-22-2010 03:50 AM
[NMR paper] Real-time NMR studies on a transient folding intermediate of barstar. Real-time NMR studies on a transient folding intermediate of barstar. http://www.ncbi.nlm.nih.gov/corehtml/query/egifs/http:--www3.interscience.wiley.com-aboutus-images-wiley_interscience_pubmed_logo_FREE_120x27.gif http://www.ncbi.nlm.nih.gov/corehtml/query/egifs/http:--www.pubmedcentral.nih.gov-corehtml-pmc-pmcgifs-pubmed-pmc.gif Related Articles Real-time NMR studies on a transient folding intermediate of barstar. Protein Sci. 1999 Jun;8(6):1286-91 Authors: Killick TR, Freund SM, Fersht AR The refolding of barstar, the intracellular...	nmrlearner	Journal club	0	08-21-2010 04:03 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