Determination of rotational correlation times of proteins in solution from carbon-13 spin-lattice relaxation measurements. Effect of magnetic field strength and anisotropic rotation

Wilbur, David J.; Norton, Raymond S.; Clouse, A. O.; Addleman, Robert; Allerhand, Adam

doi:10.1021/ja00441a059

Cited by 53 publications

(33 citation statements)

References 11 publications

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“…The rotational reorientation of the BSA molecule is anisotropic, with correlation times in the range of 25-100 ns (20,21). When the shortest correlation time (25 ns) and the theoretical analysis for isotropic reorientation are used, the predicted vi, of a methylene or methyl resonance is .100 Hz and the predicted NOE is the minimum value of 1.15 (22).…”

Section: Aqueousmentioning

confidence: 99%

Interactions of myristic acid with bovine serum albumin: a 13C NMR study.

Hamilton¹,

Cistola²,

Morrisett³

et al. 1984

Proc. Natl. Acad. Sci. U.S.A.

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Interactions of myristic acid with bovine serum albumin were studied by 13C NMR spectroscoy at 50.3 MHz using 90% isotopically substituted [1-'3C1-, [3-C] 13C]myristic acid to bovine serum albumin (from 0.7 to 5.6) revealed at least two narrow resonances for each carbon at all molar ratios. Thus, bovine serum albumin binding sites for myristic acid are heterogeneous with respect to titration behavior and with respect to the local magnetic environment at both the polar and the nonpolar ends of the fatty acid. The narrow resonances observed for the methylene and methyl carbons are inconsistent with complete immobilization of the protein-bound acid molecules. Together with spin-lattice relaxation times and nuclear Overhauser enhancements, the linewidth results indicate that bound myristic acid has internal motions that are rapid compared with overall protein tumbling and that the C-3 methylene carbon is more restricted than the terminal methyl carbon.

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Section: Aqueousmentioning

confidence: 99%

Interactions of myristic acid with bovine serum albumin: a 13C NMR study.

Hamilton¹,

Cistola²,

Morrisett³

et al. 1984

Proc. Natl. Acad. Sci. U.S.A.

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“…Several authors have reported 31p NMR measurements on DNA (3)(4)(5)(6)(7). In general, it was found that when the DNA is shorter than the persistence length, it displays a 31p linewidth of about [20][21][22][23][24][25][26][27][28][29][30] Hz and a alp T1 relaxation time near 3 sec (8-10).…”

mentioning

confidence: 99%

“…In these experiments, DNA length is known; hence TL and Ts may be fixed independently. T1(T) and T2(T) are the T, and T2 relaxation times calculated from the Wilbur (27) At high fields, 31p can experience NMR relaxation due to chemical shift anisotropy (29). Recently, however, it has been shown that at the field strengths we have used, 31P relaxation of DNA is >90% dipolar (4,5,29).…”

mentioning

confidence: 99%

Internal motions in DNA.

Hogan¹,

Jardetzky²

1979

Proc. Natl. Acad. Sci. U.S.A.

115

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We have measured the 31P and 1H NMR parameters of fractionated double-stranded DNA fragments 600, 300, and 150 base pairs long. The results indicate that, inside an intact double helix, both the deoxyribose and the sugarphosphate backbone of the helix fluctuate substantially from their equilibrium geometry. The time constant for those coupled motions is of the order of 1 nsec.The DNA structure proposed from x-ray diffraction studies is rigid. In that structure, nucleotide components of B-form DNA are locked into a 10-fold helix with base pairs oriented perpendicular to the helix axis (1).Although this DNA structure has served as an important tool in the practice of molecular biology, it is now evident that in solution B-form DNA is not rigid. The time-averaged flexibility of the DNA helix in solution approximates that of a uniform elastic wormlike coil. Recently, Barkley and Zimm have calculated the dynamic properties of the wormlike coil model (2). Their calculations predict that the average fluctuation in the twist angle between bases in DNA is about 50. The corresponding fluctuation in bending of the helix axis between bases is calculated to be about 40. Although nonexponential, both bending and twisting motions have major components with time constants between 1 and 100 nsec.NMR is a powerful tool for detecting structural fluctuations. If the motions suggested from the J3arkley-Zimm calculations involve substantial changes in DNA geometry in the time range from 1 to 100 nsec, then these motions should be detectable by both 1H and 31P NMR. Several authors have reported 31p NMR measurements on DNA (3-7). In general, it was found that when the DNA is shorter than the persistence length, it displays a 31p linewidth of about 20-30 Hz and a alp T1 relaxation time near 3 sec (8-10).Klevan et al. (4) have suggested that the 31P NMR properties (Ti, T2, nuclear Overhauser enhancement) of DNA must result from an internal motion of phosphate, independent of the overall motion of the helix. They have modeled that motion as being a rotation of backbone phosphate about C4'-C5' and P-O (w) bonds with a time constant of about 0.4 nsec. Independently, Bolton and James (5) have proposed a similar model to explain the 31P NMR properties of their DNA and RNA preparations. The two models are alike in that they consider motion within the phosphate backbone to be the only motion in the helix.Here we present a NMR study of well-defined doublestranded DNA fragments, 150-600 base pairs long. We show that, in addition to 31P NMR spectra, 1H spectra can be observed for DNA. The NMR properties of both nuclei suggest that a B-form helix experiences large fast internal motions, the best fit to the data specifying that at room temperature both the deoxyribose and the phosphate backbone of the helix fluctuate in a concerted fashion from their equilibrium geometries with a time constant near 1 nsec.The publication costs of this article were defrayed in part by page charge payment. This article must therefore be hereby marked "adverti...

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“…Large line-width may be a result of close proximity of the paramagnetic Mn2 + ion to the binding site. It is more likely that the lectin molecule, which is composed of two subunits, does not approximate a sphere in shape and thus will have a longer apparent correlation time, as was shown for bovine serum albumin ( T~ z 80 ns) [19]. Any molecular aggregation will also lead to an apparent longer correlation time.…”

Section: N M Rmentioning

confidence: 97%

Binding of [1-13C]galactose-labeled N-acetyllactosamine to Erythrina cristagalli agglutinin as studied by 13C-NMR

et al. 1985

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The equilibrium binding kinetics of enzymatically prepared N‐acetyllactosamine to the lectin from Erythrina cristagalli have been investigated by 13C‐NMR spectroscopy. Under the experimental conditions used, NMR signals in the spectrum, corresponding to both the free and bound disaccharide species, were observed for the first time. This has permitted the simultaneous determinations of the equilibrium binding constant and the number of binding sites per lectin molecule. At the relatively high lectin concentrations used (0.3–0.87 mM), the association constants determined at 31°C (∼6×103 M−1) are typically lower then those obtained by other methods employing much lower lectin concentrations. Extrapolation of the experimentally observed values to infinite dilution gave a better fit of the data (Ka∼ 1.4×104 M−1) with the binding constant determined by other methods (K∼ 1.1×104 M−1). The sugar residence time on the lectin (∼0.2 s) was determined directly from the signal's line‐width using total line‐shape analysis. Similar NMR experiments may permit an analysis of the interaction of the lectin with glycoproteins and cells labelled with 13C‐enriched galactose residues. Moreover, information on lectin‐galactose interactions at the binding site may be obtained by using galactose labeled at various carbons.

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Determination of rotational correlation times of proteins in solution from carbon-13 spin-lattice relaxation measurements. Effect of magnetic field strength and anisotropic rotation

Cited by 53 publications

References 11 publications

Interactions of myristic acid with bovine serum albumin: a 13C NMR study.

Interactions of myristic acid with bovine serum albumin: a 13C NMR study.

Internal motions in DNA.

Binding of [1-13C]galactose-labeled N-acetyllactosamine to Erythrina cristagalli agglutinin as studied by 13C-NMR

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