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The amplitude of local angular motion of purines in DNA in solution
Author(s) -
Nuutero Sirkku,
Fujimoto Bryant S.,
Flynn Peter F.,
Reid B. R.,
Ribeiro N. S.,
Schurr J. Michael
Publication year - 1994
Publication title -
biopolymers
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.556
H-Index - 125
eISSN - 1097-0282
pISSN - 0006-3525
DOI - 10.1002/bip.360340404
Subject(s) - chemistry , amplitude , molecular physics , atomic physics , guanine , nuclear magnetic resonance , analytical chemistry (journal) , optics , physics , nucleotide , biochemistry , gene , chromatography
Nuclear magnetic resonance and optical experiments are combined to determine the rms amplitude of local angular motion of purines in DNA in solution. A 12 base‐pair duplex DNA with the sequence d (CGCGAATTCGCG) 2 is deuterated at the H8 positions of adenine and guanine by exchange with solvent dklstwxzat 55°C. The deuterium nmr spectrum of this DNA is measured at 30 mg/mL at 30°C in an 11.76 Tesla magnet (76.75 MHz). The time‐resolved fluorescence polarization anisotropies (FPA) of this same sample and also a greatly diluted sample (0.215 mg/mL) were measured after addition of ethidium. FPA measurements of the dilute sample yield the hydrodynamic radius, R H = 9.94 ± 0.2 Å, while those at the nmr concentration are employed to characterize the collective motions in terms of either an enhanced viscosity or dimer formation. The rms amplitude of local angular motion was determined by analyzing the 2 H‐nmr spectrum, in particular the line width, using recently developed theory for the transverse relaxation rate (R 2 Q ) together with essential information about the collective motions from these and other optical studies. When the principal‐axis frame of the electric field gradient tensor is assumed to undergo overdamped libration around each of its three body‐fixed axes in an isotropic deflection potential, then the rms amplitude of local angular motion around any single axis is found to lie in the range 10°–11°, provided the high DNA concentration acts to enhance the viscosity, and is about 9°–11°, if it acts to produce end‐to‐end dimers. The proton nmr relaxation data of Eimer et al. are reanalyzed and shown to yield an rms amplitude of angular motion of the cytosme H5–H6 internuclear vector of 9°–10°, depending upon its orientation with respect to the helix axis. In all of these analyses, full account is taken of the collective twisting and bending deformations, which have a small but significant effect on the results. It is shown that the rms amplitudes of local angular motion do not depend strongly on the model (potential), provided that isotropic rotation around the same number of axes is allowed and that one compares rms angles of the same dimensionality. The rms amplitudes of local angular motion in solution are comparable to those observed for the same sequence at low levels of hydration in the solid state. © 1994 John Wiley & Sons, Inc.