Related ArticlesCharacterization of the internal motions of a chimeric protein by 13C NMR highlights the important dynamic consequences of the engineering on a millisecond time scale.
Eur J Biochem. 2000 Nov;267(22):6519-33
Authors: Wolff N, Guenneugues M, Gilquin B, Drakopoulou E, Vita C, Ménez A, Zinn-Justin S
By transferring the central curaremimetic beta hairpin of the snake toxin alpha into the scaffold of the scorpion charybdotoxin, a chimeric protein was constructed that reproduced the three-dimensional structure and partially reproduced the function of the parent beta hairpin, without perturbing the three-dimensional structure of the scaffold [1]. Picosecond to hour time scale motions of charybdotoxin and the engineered protein were observed, in order to evaluate the dynamic consequences of the six deletions and eight mutations differentiating the two molecules. The chimeric protein dynamics were also compared to that of toxin alpha, in order to examine the beta hairpin motions in both structural contexts. Thus, 13C R1, R1rho and 1H-->13C nOe were measured for all the CalphaHalpha and threonine CbetaHbeta vectors. As the proteins were not labeled, accordion techniques combined to coherence selection by pulsed field gradients and preservation of magnetization following equivalent pathways were used to considerably reduce the spectrometer time needed. On one hand, we observed that the chimeric protein and charybdotoxin are subjected to similar picosecond to nanosecond time scale motions except around the modified beta sheet region. The chimeric protein also exhibits an additional millisecond time scale motion on its whole sequence, and its beta structure is less stable on a minute to hour time scale. On the other hand, when the beta hairpin dynamics is compared in two different structural contexts, i.e. in the chimeric protein and the curaremimetic toxin alpha, the picosecond to nanosecond time scale motions are fairly conserved. However, the microsecond to millisecond time scale motions are different on most of the beta hairpin sequence, and the beta sheet seems more stable in toxin alpha than in the chimera. The slower microsecond to hour time scale motions seem to be extremely sensitive to the structural context, and thus poorly transferred from one protein to another.
[NMR structure and dynamics of the chimeric protein SH3-F2].
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Mol Biol (Mosk). 2010 Nov-Dec;44(6):1064-74
Authors:
For the further elucidation of structural and dynamic principles of protein self-organization and protein-ligand interactions the design of new chimeric protein SH3-F2 was made and genetically engineered construct was created. The SH3-F2 amino acid sequence consists of polyproline ligand mgAPPLPPYSA, GG linker and the sequence of spectrin SH3 domain circular permutant S19-P20s. Structural and dynamics properties of the protein were studied by high-resolution NMR. According to NMR data the...
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Can Enzyme Engineering Benefit from the Modulation of Protein Motions? Lessons Learned from NMR Relaxation Dispersion Experiments.
Can Enzyme Engineering Benefit from the Modulation of Protein Motions? Lessons Learned from NMR Relaxation Dispersion Experiments.
Can Enzyme Engineering Benefit from the Modulation of Protein Motions? Lessons Learned from NMR Relaxation Dispersion Experiments.
Protein Pept Lett. 2011 Jan 11;
Authors:
Despite impressive progress in protein engineering and design, our ability to create new and efficient enzyme activities remains a laborious and time-consuming endeavor. In the past few years, intricate combinations of rational mutagenesis, directed...
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[NMR paper] The effects of mutations on motions of side-chains in protein L studied by 2H NMR dyn
The effects of mutations on motions of side-chains in protein L studied by 2H NMR dynamics and scalar couplings.
Related Articles The effects of mutations on motions of side-chains in protein L studied by 2H NMR dynamics and scalar couplings.
J Mol Biol. 2003 Jun 6;329(3):551-63
Authors: Millet O, Mittermaier A, Baker D, Kay LE
Recently developed 2H spin relaxation experiments are applied to study the dynamics of methyl-containing side-chains in the B1 domain of protein L and in a pair of point mutants of the domain, F22L and A20V. X-ray and...
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[NMR paper] Backbone motions in a crystalline protein from field-dependent 2H-NMR relaxation and
Backbone motions in a crystalline protein from field-dependent 2H-NMR relaxation and line-shape analysis.
Related Articles Backbone motions in a crystalline protein from field-dependent 2H-NMR relaxation and line-shape analysis.
Biopolymers. 2000 Jan;53(1):9-18
Authors: Mack JW, Usha MG, Long J, Griffin RG, Wittebort RJ
We have used 2H-nmr to study backbone dynamics of the 2H-labeled, slowly exchanging amide sites of fully hydrated, crystalline hen egg white lysozyme. Order parameters are determined from the residual quadrupole coupling and...
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11-18-2010 09:15 PM
Comparison of NMR and crystal structures highlights conformational isomerism in prote
Comparison of NMR and crystal structures highlights conformational isomerism in protein active sites.
Related Articles Comparison of NMR and crystal structures highlights conformational isomerism in protein active sites.
Acta Crystallogr Sect F Struct Biol Cryst Commun. 2010 Oct 1;66(Pt 10):1393-405
Authors: Serrano P, Pedrini B, Geralt M, Jaudzems K, Mohanty B, Horst R, Herrmann T, Elsliger MA, Wilson IA, Wüthrich K
The JCSG has recently developed a protocol for systematic comparisons of high-quality crystal and NMR structures of proteins. In this...
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[NMR paper] Contributions to protein entropy and heat capacity from bond vector motions measured
Contributions to protein entropy and heat capacity from bond vector motions measured by NMR spin relaxation.
http://www.ncbi.nlm.nih.gov/corehtml/query/egifs/http:--linkinghub.elsevier.com-ihub-images-PubMedLink.gif Related Articles Contributions to protein entropy and heat capacity from bond vector motions measured by NMR spin relaxation.
J Mol Biol. 1997 Oct 10;272(5):790-804
Authors: Yang D, Mok YK, Forman-Kay JD, Farrow NA, Kay LE
The backbone dynamics of both folded and unfolded states of staphylococcal nuclease (SNase) and the N-terminal...
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[NMR paper] Internal motions of apo-neocarzinostatin as studied by 13C NMR methine relaxation at
Internal motions of apo-neocarzinostatin as studied by 13C NMR methine relaxation at natural abundance.
Related Articles Internal motions of apo-neocarzinostatin as studied by 13C NMR methine relaxation at natural abundance.
J Biomol NMR. 1995 Apr;5(3):233-44
Authors: Mispelter J, Lefèvre C, Adjadj E, Quiniou E, Favaudon V
Dynamics of the backbone and some side chains of apo-neocarzinostatin, a 10.7 kDa carrier protein, have been studied from 13C relaxation rates R1, R2 and steady-state 13C-(1H) NOEs, measured at natural abundance. Relaxation...
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[NMR paper] Effect of antibody binding on protein motions studied by hydrogen-exchange labeling a
Effect of antibody binding on protein motions studied by hydrogen-exchange labeling and two-dimensional NMR.
Related Articles Effect of antibody binding on protein motions studied by hydrogen-exchange labeling and two-dimensional NMR.
Biochemistry. 1992 Nov 10;31(44):10678-85
Authors: Mayne L, Paterson Y, Cerasoli D, Englander SW
We have used hydrogen-exchange labeling detected by 2D NMR to study antibody-protein interactions for two monoclonal antibodies raised against horse cytochrome c. The data show that these antibodies bind mainly to the...