[NMR paper] Effects of Phe-to-Trp mutation and fluorotryptophan incorporation on the solution structure of cardiac troponin C, and analysis of its suitability as a potential probe for in situ NMR studies.
Effects of Phe-to-Trp mutation and fluorotryptophan incorporation on the solution structure of cardiac troponin C, and analysis of its suitability as a potential probe for in situ NMR studies.
Related ArticlesEffects of Phe-to-Trp mutation and fluorotryptophan incorporation on the solution structure of cardiac troponin C, and analysis of its suitability as a potential probe for in situ NMR studies.
Protein Sci. 2005 Sep;14(9):2447-60
Authors: Wang X, Mercier P, Letourneau PJ, Sykes BD
19F NMR spectroscopy is potentially a powerful tool for probing protein properties in situ. However, results obtained using this technique are relevant only if the 19F probe offers minimal perturbation to the surrounding environment. In this paper, we examine the effect of 5-fluorotryptophan (5fW) incorporation on the three-dimensional structure of cardiac troponin-C (cTnC), with the intention of developing a 19F-labeled TnC for use in in situ 19FNMR. We find that, in general, 5fW does not perturb the structure of the protein significantly. Replacement of residue Phe 153 with 5fW produces no noticeable change in protein conformation. However, replacement of residue Phe 104 with 5fW produces a folding behavior that is dependent on the Escherichia coli strain used to express the mutant. The orientations of the indole rings in these mutants are such that the Trp residue adopts a chi2 of approximately 90 degrees in the F104W mutant and approximately -100 degrees in the F153W mutant. Using results from 19F-1H heteronuclear NOE experiment, we show the replacement of L-Trp with 5fW at these positions does not change the orientation of the indole ring and the spread of the 5fW side-chain dihedral angles increases moderately for the F104(5fW) mutant and not at all for the F153(5fW) mutant. Based on these structures, we conclude that the substitution of Phe by 5fW at these two positions has minimal effects on the structure of cTnC and that the 5fW indole rings in both mutants have well defined orientation, making the two mutants viable candidates for use in in situ 19F NMR spectroscopy.
[NMR paper] The interaction of the bisphosphorylated N-terminal arm of cardiac troponin I-A 31P-N
The interaction of the bisphosphorylated N-terminal arm of cardiac troponin I-A 31P-NMR study.
Related Articles The interaction of the bisphosphorylated N-terminal arm of cardiac troponin I-A 31P-NMR study.
FEBS Lett. 2002 Feb 27;513(2-3):289-93
Authors: Schmidtmann A, Lohmann K, Jaquet K
Cardiac troponin I, the inhibitory subunit of the heterotrimeric cardiac troponin (cTn) complex is phosphorylated by protein kinase A at two serine residues located in its heart-specific N-terminal extension. This flexible arm interacts at different sites...
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[NMR paper] 19F NMR relaxation studies on 5-fluorotryptophan- and tetradeutero-5-fluorotryptophan
19F NMR relaxation studies on 5-fluorotryptophan- and tetradeutero-5-fluorotryptophan-labeled E. coli glucose/galactose receptor.
Related Articles 19F NMR relaxation studies on 5-fluorotryptophan- and tetradeutero-5-fluorotryptophan-labeled E. coli glucose/galactose receptor.
J Biomol NMR. 1996 Jun;7(4):261-72
Authors: Luck LA, Vance JE, O'Connell TM, London RE
19F NMR relaxation studies have been carried out on a fluorotryptophan-labeled E. coli periplasmic glucose/galactose receptor (GGR). The protein was derived from E. coli grown on a...
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[NMR paper] Cardiac troponin I induced conformational changes in cardiac troponin C as monitored
Cardiac troponin I induced conformational changes in cardiac troponin C as monitored by NMR using site-directed spin and isotope labeling.
Related Articles Cardiac troponin I induced conformational changes in cardiac troponin C as monitored by NMR using site-directed spin and isotope labeling.
Biochemistry. 1995 Oct 17;34(41):13343-52
Authors: Kleerekoper Q, Howarth JW, Guo X, Solaro RJ, Rosevear PR
Conformational changes in both free cardiac troponin C (cTnC) and in complex with a recombinant troponin I protein were observed by means of a...
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[NMR paper] NMR studies delineating spatial relationships within the cardiac troponin I-troponin
NMR studies delineating spatial relationships within the cardiac troponin I-troponin C complex.
Related Articles NMR studies delineating spatial relationships within the cardiac troponin I-troponin C complex.
J Biol Chem. 1994 Sep 23;269(38):23731-5
Authors: Krudy GA, Kleerekoper Q, Guo X, Howarth JW, Solaro RJ, Rosevear PR
NMR spectroscopy and selective isotope labeling of both recombinant cardiac troponin C (cTnC3) and a truncated cardiac troponin I (cTnI/NH2) lacking the N-terminal 32-amino acid cardiac-specific sequence have been used to...
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[NMR paper] Comparative NMR studies on cardiac troponin C and a mutant incapable of binding calci
Comparative NMR studies on cardiac troponin C and a mutant incapable of binding calcium at site II.
Related Articles Comparative NMR studies on cardiac troponin C and a mutant incapable of binding calcium at site II.
Biochemistry. 1991 Oct 22;30(42):10236-45
Authors: Brito RM, Putkey JA, Strynadka NC, James MN, Rosevear PR
One- and two-dimensional NMR techniques were used to study both the influence of mutations on the structure of recombinant normal cardiac troponin C (cTnC3) and the conformational changes induced by Ca2+ binding to site II,...
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08-21-2010 11:12 PM
[NMR paper] Comparative NMR studies on cardiac troponin C and a mutant incapable of binding calci
Comparative NMR studies on cardiac troponin C and a mutant incapable of binding calcium at site II.
Related Articles Comparative NMR studies on cardiac troponin C and a mutant incapable of binding calcium at site II.
Biochemistry. 1991 Oct 22;30(42):10236-45
Authors: Brito RM, Putkey JA, Strynadka NC, James MN, Rosevear PR
One- and two-dimensional NMR techniques were used to study both the influence of mutations on the structure of recombinant normal cardiac troponin C (cTnC3) and the conformational changes induced by Ca2+ binding to site II,...
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[NMR paper] Sites phosphorylated in bovine cardiac troponin T and I. Characterization by 31P-NMR
Sites phosphorylated in bovine cardiac troponin T and I. Characterization by 31P-NMR spectroscopy and phosphorylation by protein kinases.
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 Sites phosphorylated in bovine cardiac troponin T and I. Characterization by 31P-NMR spectroscopy and phosphorylation by protein kinases.
Eur J Biochem. 1990 Jul 5;190(3):575-82
Authors: Swiderek K, Jaquet K, Meyer HE, Schächtele C, Hofmann F, Heilmeyer LM
...
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[NMR paper] NMR analysis of cardiac troponin C-troponin I complexes: effects of phosphorylation.
NMR analysis of cardiac troponin C-troponin I complexes: effects of phosphorylation.
http://www.ncbi.nlm.nih.gov/corehtml/query/egifs/http:--linkinghub.elsevier.com-ihub-images-PubMedLink.gif Related Articles NMR analysis of cardiac troponin C-troponin I complexes: effects of phosphorylation.
FEBS Lett. 1999 Jun 18;453(1-2):107-12
Authors: Finley N, Abbott MB, Abusamhadneh E, Gaponenko V, Dong W, Gasmi-Seabrook G, Howarth JW, Rance M, Solaro RJ, Cheung HC, Rosevear PR
Phosphorylation of the cardiac specific amino-terminus of troponin I has...
Effects of Phe-to-Trp mutation and fluorotryptophan incorporation on the solution structure of cardiac troponin C, and analysis of its suitability as a potential probe for in situ NMR studies.
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