NMR paper Lead activation of protein kinase C from rat brain. Determination of free calcium, le

		BioNMR > Educational resources > Journal club
[NMR paper] Lead activation of protein kinase C from rat brain. Determination of free calcium, le

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

08-22-2010, 03:33 AM

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

Lead activation of protein kinase C from rat brain. Determination of free calcium, le

Lead activation of protein kinase C from rat brain. Determination of free calcium, lead, and zinc by 19F NMR.

Related Articles Lead activation of protein kinase C from rat brain. Determination of free calcium, lead, and zinc by 19F NMR.

J Biol Chem. 1994 Jan 14;269(2):834-7

Authors: Long GJ, Rosen JF, Schanne FA

Lead (Pb2+) has been reported to activate calcium/phospholipid-dependent protein kinase C (PKC) at subnanomolar concentrations (Markovac, J., and Goldstein, G. W. (1988) Nature 334, 732-734); however, others have failed to find any Pb(2+)-induced activation of PKC (Murakami, K., Feng, G., and Chen, S. G. (1993) J. Pharmacol. Exp. Ther. 264, 757-761). In neither of these studies was the actual free Pb2+ or Ca2+ concentration measured. In this study, 1,2-bis(2-amino-5-fluorophenoxy)ethane N,N,N',N'-tetraacetic acid (5F-BAPTA) was used to buffer Pb2+ and Ca2+ concentrations in the PKC reaction mixture. The specific free ion concentrations of Pb2+ and Ca2+, as well as Zn2+ and other divalent cations contained in the PKC reaction mixtures, were determined by 19F NMR spectroscopy. Using this approach to set and confirm the free Pb2+ and Ca2+ concentrations, we measured the Pb(2+)-dependent and the Ca(2+)-dependent activation of phosphotydylserine/diolein-dependent incorporation of 32P from ATP into histone and endogenous acid precipitable proteins in the 100,000 x g supernatant from homogenized rat brain cortex. We found that free Pb2+ activates PKC in the range from 10(-11) to 10(-8) M, Kact = 5.5 x 10(-11) M, while Ca2+ activates PKC in the range from 10(-8) to 10(-5) M, Kact = 2.56 x 10(-7) M. These findings clearly resolve the activation of PKC by subnanomolar concentrations of free Pb2+ from activation induced by Ca2+ or other divalent cations. Furthermore, it documents the utility of 5F-BAPTA as buffer and indicator when resolving the contributions of multiple divalent cations in biochemical processes.

PMID: 8288636 [PubMed - indexed for MEDLINE]

Source: PubMed

Did you find this post helpful?

Similar Threads
Thread	Thread Starter	Forum	Replies	Last Post
Fast methionine-based solution structure determination of calcium-calmodulin complexes Fast methionine-based solution structure determination of calcium-calmodulin complexes Abstract Here we present a novel NMR method for the structure determination of calcium-calmodulin (Ca2+-CaM)-peptide complexes from a limited set of experimental restraints. A comparison of solved CaM-peptide structures reveals invariability in CaMâ??s backbone conformation and a structural plasticity in CaMâ??s domain orientation enabled by a flexible linker. Knowing this, the collection and analysis of an extensive set of NOESY spectra is redundant. Although RDCs can define CaM domain orientation in...	nmrlearner	Journal club	0	03-03-2011 02:06 AM
[NMR paper] Rapid and accurate structure determination of coiled-coil domains using NMR dipolar couplings: application to cGMP-dependent protein kinase Ialpha. Rapid and accurate structure determination of coiled-coil domains using NMR dipolar couplings: application to cGMP-dependent protein kinase Ialpha. Related Articles Rapid and accurate structure determination of coiled-coil domains using NMR dipolar couplings: application to cGMP-dependent protein kinase Ialpha. Protein Sci. 2005 Sep;14(9):2421-8 Authors: Schnell JR, Zhou GP, Zweckstetter M, Rigby AC, Chou JJ Coiled-coil motifs play essential roles in protein assembly and molecular recognition, and are therefore the targets of many ongoing...	nmrlearner	Journal club	0	12-01-2010 06:56 PM
[NMR paper] Rotational dynamics of calcium-free calmodulin studied by 15N-NMR relaxation measurem Rotational dynamics of calcium-free calmodulin studied by 15N-NMR relaxation measurements. http://www.ncbi.nlm.nih.gov/corehtml/query/egifs/http:--www3.interscience.wiley.com-aboutus-images-wiley_interscience_pubmed_logo_FREE_120x27.gif Related Articles Rotational dynamics of calcium-free calmodulin studied by 15N-NMR relaxation measurements. Eur J Biochem. 1995 Jun 15;230(3):1014-24 Authors: Tjandra N, Kuboniwa H, Ren H, Bax A The backbone motions of calcium-free Xenopus calmodulin have been characterized by measurements of the 15N...	nmrlearner	Journal club	0	08-22-2010 03:41 AM
[NMR paper] Lead activation of protein kinase C from rat brain. Determination of free calcium, le Lead activation of protein kinase C from rat brain. Determination of free calcium, lead, and zinc by 19F NMR. Related Articles Lead activation of protein kinase C from rat brain. Determination of free calcium, lead, and zinc by 19F NMR. J Biol Chem. 1994 Jan 14;269(2):834-7 Authors: Long GJ, Rosen JF, Schanne FA Lead (Pb2+) has been reported to activate calcium/phospholipid-dependent protein kinase C (PKC) at subnanomolar concentrations (Markovac, J., and Goldstein, G. W. (1988) Nature 334, 732-734); however, others have failed to find any...	nmrlearner	Journal club	0	08-22-2010 03:33 AM
[NMR paper] Activation of the phosphosignaling protein CheY. I. Analysis of the phosphorylated co Activation of the phosphosignaling protein CheY. I. Analysis of the phosphorylated conformation by 19F NMR and protein engineering. http://www.ncbi.nlm.nih.gov/corehtml/query/egifs/http:--www.pubmedcentral.nih.gov-corehtml-pmc-pmcgifs-pubmed-pmc-MS.gif Related Articles Activation of the phosphosignaling protein CheY. I. Analysis of the phosphorylated conformation by 19F NMR and protein engineering. J Biol Chem. 1993 Jun 25;268(18):13081-8 Authors: Drake SK, Bourret RB, Luck LA, Simon MI, Falke JJ CheY, the 14-kDa response regulator protein of...	nmrlearner	Journal club	0	08-21-2010 11:53 PM
[NMR paper] Determination of the solution structure of a synthetic two-site calcium-binding homod Determination of the solution structure of a synthetic two-site calcium-binding homodimeric protein domain by NMR spectroscopy. Related Articles Determination of the solution structure of a synthetic two-site calcium-binding homodimeric protein domain by NMR spectroscopy. Biochemistry. 1992 Oct 13;31(40):9572-80 Authors: Shaw GS, Hodges RS, Sykes BD The solution structure of a 34-residue synthetic calcium-binding peptide from site III of chicken troponin-C has been determined by 1H NMR spectroscopy. In solution and in the presence of calcium...	nmrlearner	Journal club	0	08-21-2010 11:45 PM
[NMR paper] 19F-NMR study of the effect of lead on intracellular free calcium in human platelets. 19F-NMR study of the effect of lead on intracellular free calcium in human platelets. http://www.ncbi.nlm.nih.gov/corehtml/query/egifs/http:--linkinghub.elsevier.com-ihub-images-PubMedLink.gif Related Articles 19F-NMR study of the effect of lead on intracellular free calcium in human platelets. Biochim Biophys Acta. 1991 May 17;1092(3):341-6 Authors: Dowd TL, Gupta RK Lead has been shown to affect calcium homeostasis. However, there is no prior evidence to indicate an effect of low concentrations of lead in the environment (approximately 1...	nmrlearner	Journal club	0	08-21-2010 11:16 PM
[NMR paper] NMR detection of creatine kinase expressed in liver of transgenic mice: determination NMR detection of creatine kinase expressed in liver of transgenic mice: determination of free ADP levels. http://www.ncbi.nlm.nih.gov/corehtml/query/egifs/http:--www.pubmedcentral.nih.gov-corehtml-pmc-pmcgifs-pubmed-pmc.gif Related Articles NMR detection of creatine kinase expressed in liver of transgenic mice: determination of free ADP levels. Proc Natl Acad Sci U S A. 1990 Apr;87(8):3112-6 Authors: Koretsky AP, Brosnan MJ, Chen LH, Chen JD, Van Dyke T To use the equilibrium established by creatine kinase (CK) to determine hepatic free ADP...	nmrlearner	Journal club	0	08-21-2010 10:48 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