Related ArticlesCharacterization of trimethyllysine 115 in calmodulin by 14N and 13C NMR spectroscopy.
J Biol Chem. 1994 Feb 18;269(7):5099-105
Authors: Zhang M, Huque E, Vogel HJ
In this paper, we describe three approaches to study the single trimethyllysine 115 in calmodulin. First, 14N NMR spectroscopy has been used as a novel spectroscopic tool. Because of the unique symmetrical tetrahedral substitution of its side chain, the trimethyllysine residue gives rise to a sharp 14N NMR resonance; hence, this has allowed the detection and quantitation of the level of trimethylation. Trimethyllysine side chains of bovine testis calmodulin and yeast cytochrome c were shown to have a high mobility in aqueous solution as determined by 14N NMR relaxation measurements. Second, we have purified mammalian calmodulin from an overproducing Escherichia coli strain. By comparison of the 1H-13C heteronuclear multiple quantum coherence spectra of 13C-dimethylated calmodulin samples from bovine testis and E. coli, the resonance for Lys-115 in bacterially expressed calmodulin could be identified. pH titration experiments showed that epsilon-NH2 group of Lys-115 has a normal pKa value both in the apo and Ca2+ forms of the protein and in a complex of calmodulin with a 22-residue calmodulin-binding peptide derived from myosin light chain kinase. Third, we have shown that mutation of Lys-115 to the uncharged Gln residue does not alter the ability of the protein to stimulate the enzymes cyclic nucleotide phosphodiesterase and myosin light chain kinase. These results show that the trimethylation of Lys-115 is not caused by an unusual pKa and reactivity of its epsilon-NH2 group and that its side chain remains flexible. Moreover, our data suggest that the introduction of a permanent positive charge on Lys-115 by trimethylation is also not the major reason for this specific post-translational modification.
Human multiprotein bridging factor 1 and Calmodulin do not interact in vitro as confirmed by NMR spectroscopy and CaM-agarose affinity chromatography.
Human multiprotein bridging factor 1 and Calmodulin do not interact in vitro as confirmed by NMR spectroscopy and CaM-agarose affinity chromatography.
Human multiprotein bridging factor 1 and Calmodulin do not interact in vitro as confirmed by NMR spectroscopy and CaM-agarose affinity chromatography.
Protein Expr Purif. 2011 Jul 14;
Authors: Babini E, Hu X, Parigi G, Vignali M
The human multiprotein bridging factor 1 (hMBF1) has been established in different cellular types to have the role of transcriptional coactivator. It is also reported to be...
nmrlearner
Journal club
0
07-26-2011 09:30 PM
The calponin regulatory region is intrinsically unstructured: novel insight into actin-calponin and calmodulin-calponin interfaces using NMR spectroscopy.
The calponin regulatory region is intrinsically unstructured: novel insight into actin-calponin and calmodulin-calponin interfaces using NMR spectroscopy.
The calponin regulatory region is intrinsically unstructured: novel insight into actin-calponin and calmodulin-calponin interfaces using NMR spectroscopy.
Biophys J. 2011 Apr 6;100(7):1718-28
Authors: Pfuhl M, Al-Sarayreh S, El-Mezgueldi M
Calponin is an actin- and calmodulin-binding protein believed to regulate the function of actin. Low-resolution studies based on proteolysis established that...
nmrlearner
Journal club
0
04-06-2011 10:54 AM
[NMR paper] Functional characterization and NMR spectroscopy on full-length Vpu from HIV-1 prepar
Functional characterization and NMR spectroscopy on full-length Vpu from HIV-1 prepared by total chemical synthesis.
Related Articles Functional characterization and NMR spectroscopy on full-length Vpu from HIV-1 prepared by total chemical synthesis.
J Am Chem Soc. 2004 Mar 3;126(8):2439-46
Authors: Kochendoerfer GG, Jones DH, Lee S, Oblatt-Montal M, Opella SJ, Montal M
Vpu is an 81-residue integral membrane protein encoded in the HIV-1 genome that is of considerable interest because it plays important roles in the release of virus particles...
nmrlearner
Journal club
0
11-24-2010 09:25 PM
[NMR paper] Preparation, characterization, and NMR spectroscopy of encapsulated proteins dissolve
Preparation, characterization, and NMR spectroscopy of encapsulated proteins dissolved in low viscosity fluids.
Related Articles Preparation, characterization, and NMR spectroscopy of encapsulated proteins dissolved in low viscosity fluids.
J Biomol NMR. 2003 Apr;25(4):313-23
Authors: Babu CR, Flynn PF, Wand AJ
Encapsulating a protein in a reverse micelle and dissolving it in a low-viscosity solvent can lower the rotational correlation time of a protein and thereby provides a novel strategy for studying proteins in a variety of contexts. The...
nmrlearner
Journal club
0
11-24-2010 09:01 PM
[NMR paper] Calcium-induced refolding of the calmodulin V136G mutant studied by NMR spectroscopy:
Calcium-induced refolding of the calmodulin V136G mutant studied by NMR spectroscopy: evidence for interaction between the two globular domains.
Related Articles Calcium-induced refolding of the calmodulin V136G mutant studied by NMR spectroscopy: evidence for interaction between the two globular domains.
Biochemistry. 2000 Dec 26;39(51):15920-31
Authors: Fefeu S, Biekofsky RR, McCormick JE, Martin SR, Bayley PM, Feeney J
The Ca(2+) titration of the (15)N-labeled mutant V136G calmodulin has been monitored using (1)H-(15)N HSQC NMR spectra. Up...
nmrlearner
Journal club
0
11-19-2010 08:29 PM
[NMR paper] Recombinant locust apolipophorin III: characterization and NMR spectroscopy.
Recombinant locust apolipophorin III: characterization and NMR spectroscopy.
Related Articles Recombinant locust apolipophorin III: characterization and NMR spectroscopy.
Biochim Biophys Acta. 1998 Jul 31;1393(1):99-107
Authors: Weers PM, Wang J, Van der Horst DJ, Kay CM, Sykes BD, Ryan RO
Apolipophorin III (apoLp-III) from the locust Locusta migratoria is an exchangeable apolipoprotein that reversibly binds to lipoproteins. During lipid binding the protein has been proposed to undergo a major conformational change. To study the mechanism of...
nmrlearner
Journal club
0
11-17-2010 11:15 PM
Characterization of caged compounds binding to proteins by NMR spectroscopy.
Characterization of caged compounds binding to proteins by NMR spectroscopy.
Characterization of caged compounds binding to proteins by NMR spectroscopy.
Biochem Biophys Res Commun. 2010 Aug 27;
Authors: Bandorowicz-Pikula J, Buchet R, Cañada FJ, Clémancey M, Groves P, Jiménez-Barbero J, Lancelin JM, Marcillat O, Pikula S, Sekrecka-Belniak A, Strzelecka-Kiliszek A
Photolysable caged ligands are used to investigate protein function and activity. Here, we investigate the binding properties of caged nucleotides and their photo released...
nmrlearner
Journal club
0
09-02-2010 03:58 PM
[NMR paper] Characterization of trimethyllysine 115 in calmodulin by 14N and 13C NMR spectroscopy
Characterization of trimethyllysine 115 in calmodulin by 14N and 13C NMR spectroscopy.
Related Articles Characterization of trimethyllysine 115 in calmodulin by 14N and 13C NMR spectroscopy.
J Biol Chem. 1994 Feb 18;269(7):5099-105
Authors: Zhang M, Huque E, Vogel HJ
In this paper, we describe three approaches to study the single trimethyllysine 115 in calmodulin. First, 14N NMR spectroscopy has been used as a novel spectroscopic tool. Because of the unique symmetrical tetrahedral substitution of its side chain, the trimethyllysine residue...