1H,13C and15N resonance assignment for barnase

Korzhnev, Dmitry M.; Bocharov, Eduard V.; Zhuravlyova, A. V.; Tischenko, E. V.; Reibarkh, Mikhail; Ermolyuk, Yaroslav S.; Schulga, Alexey; Kirpichnikov, Mikhaǐl P.; Billeter, Martin; Arseniev, Alexander S.

doi:10.1007/bf03162451

Cited by 7 publications

(7 citation statements)

References 12 publications

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“…45 A barnasebarstar complex was generated by adding aliquots from a 12 mM solution of purified barstar to a 1.3 mM barnase solution until monomer signals disappeared in 1 H-15 N 2D heteronuclear single quantum coherence spectra, which resulted in a final barnase to barstar molar ratio of 1:1 and a final complex concentration of 1.17 mM (based on amount of barstar added). A fraction of this sample was diluted to produce another sample of the barnase-barstar complex with a concentration of 0.17 mM.…”

Section: Nmr Sample Preparationmentioning

confidence: 99%

Propagation of Dynamic Changes in Barnase Upon Binding of Barstar: An NMR and Computational Study

Zhuravleva

Korzhnev

Nolde

et al. 2007

Journal of Molecular Biology

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Section: Nmr Sample Preparationmentioning

confidence: 99%

Propagation of Dynamic Changes in Barnase Upon Binding of Barstar: An NMR and Computational Study

Zhuravleva

Korzhnev

Nolde

et al. 2007

Journal of Molecular Biology

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“…Data were processed using FELIX (Accelrys, San Diego, CA), and peaks were analyzed using Excel (Microsoft, Seattle, WA). Chemical shift assignments (14) were confirmed using standard triple resonance NMR experiments. For the 13 C HSQC spectra, folding was used in the indirect dimension, which means that there was some overlapping of side-chain signals.…”

Section: Nmr Experimentsmentioning

confidence: 99%

“…Crystal structures of the free enzyme and of complexes with several nucleotide inhibitors, as well as its physiological partner barstar, show that the structure changes very little on binding to substrate, but suggest that there is a loss of mobility in some regions, particularly in the active site (12,13). Barnase has been extensively studied by NMR, and resonance assignments (14) as well as 15 N relaxation parameters (15) are available. It shows substrate specificity for cleavage after guanine bases, and the binding site has a subsite where guanine binds, both in the crystal (13) and in solution (16).…”

Section: Introductionmentioning

confidence: 99%

Pressure-Dependent Structure Changes in Barnase on Ligand Binding Reveal Intermediate Rate Fluctuations

Wilton

Kitahara

Akasaka

et al. 2009

Biophysical Journal

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In this work we measured 1H NMR chemical shifts for the ribonuclease barnase at pressures from 3 MPa to 200 MPa, both free and bound to d(CGAC). Shift changes with pressure were used as restraints to determine the change in structure with pressure. Free barnase is compressed by approximately 0.7%. The largest changes are on the ligand-binding face close to Lys-27, which is the recognition site for the cleaved phosphate bond. This part of the protein also contains the buried water molecules. In the presence of d(CGAC), the compressibility is reduced by approximately 70% and the region of structural change is altered: the ligand-binding face is now almost incompressible, whereas changes occur at the opposite face. Because compressibility is proportional to mean square volume fluctuation, we conclude that in free barnase, volume fluctuation is largest close to the active site, but when the inhibitor is bound, the fluctuations become much smaller and are located mainly on the opposite face. The timescale of the fluctuations is nanoseconds to microseconds, consistent with the degree of ordering required for the fluctuations, which are intermediate between rapid uncorrelated side-chain dynamics and slow conformational transitions. The high-pressure technique is therefore useful for characterizing motions on this relatively inaccessible timescale.

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“…PR promoter. Recently the analog of this expression system was successfully used for the biosynthesis of ribonuclease barnase enriched by ZH, 13C and ~SN isotopes [9]. An overnight culture of Escherichia coli BL21 cells harboring the neurotoxin expression vector was grown at 28~ in minimal medium.…”

Section: Biosynthesis Of Lscflsn Uniformly Labeled Neurotoxin Hmentioning

confidence: 99%

Resonance assignment of13C−15N-labeled snake neurotoxin II fromNaja oxiana

et al. 2003

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Neurotoxin II from Naja oxiana venom is a short-chain snake curaremimetic neurotoxin containing four disulfide bonds. We obtained ~3C-~SN-labeled neurotoxin II to study its intemal dynamics and surface properties with atomic resolution. The recombinant protein has the native spatial structure and is biologically active. The nearly complete assignment of ~H, ~3C and ~SN resonances for neurotoxin II was obtained by heteronuclear triple-resonance nuclear magnetic resonance spectroscopy. Analysis of the secondary chemical shifts of the ~H ~, ~3C~, ~3C~ and ~3CO nuclei reveal their strong correlation with the protein secondary structure.

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1H,13C and15N resonance assignment for barnase

Cited by 7 publications

References 12 publications

Propagation of Dynamic Changes in Barnase Upon Binding of Barstar: An NMR and Computational Study

Propagation of Dynamic Changes in Barnase Upon Binding of Barstar: An NMR and Computational Study

Pressure-Dependent Structure Changes in Barnase on Ligand Binding Reveal Intermediate Rate Fluctuations

Resonance assignment of13C−15N-labeled snake neurotoxin II fromNaja oxiana

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