RESUMO
Anomalous electric birefringence signals of a sonicated and column-fractionated medium-size calf thymus DNA sample (bp=570) in Na(+) solutions were measured at 7 degrees C. The reversing-pulse electric birefringence (RPEB) signal pattern was theoretically calculated in the low electric field region for two axially symmetric models coexisting in equilibrium in solution. The RPEB theory is based on the electric dipole moment due to ion-fluctuation along the longitudinal direction and the electric polarizability anisotropy (Deltaalpha'), together with various electric and optical parameters assigned to the models. An analytical method was developed for the steady-state birefringence of the two-component system in a wide range of electric fields. The NaDNA samples exhibit complex RPEB patterns mixed with negative- and positive-going profiles. An experimental RPEB signal of NaDNA at an absorbance (A(260)) of 8 was fitted to theoretical curve at weak electric fields. The anomalous RPEB signal was attributed to the component 2, which shows a dip in the buildup and another in the reverse processes with a positive sign and a larger relaxation time. For the component 1, a normal DNA profile with negative sign is associated with a narrow dip in the reverse and a faster relaxation time in the decay signal. The field-strength dependence of observed steady-state birefringence delta(infinity) could be fitted for NaDNA at A(260)=8 by the SUSID orientation function with saturated ionic and electronic moments. An apparent positive maximum and the sign reversal in delta(infinity) at weak electric fields is an interplay between the positive component 2 with positive optical factor Deltag and negative Deltaalpha' and the negative component 1 with negative Deltag and positive Deltaalpha'. Possible conformation of two DNA components involved in solution was estimated.
Assuntos
DNA/química , Eletricidade , Eletroquímica/métodos , Animais , Anisotropia , Birrefringência , Bovinos , Cromatografia em Gel , Diálise , Cinética , Luz , Modelos Teóricos , Peso Molecular , Conformação de Ácido Nucleico , Concentração Osmolar , Espalhamento de Radiação , Cloreto de Sódio/química , Soluções , Sonicação , Espectrofotometria Ultravioleta , Timo/químicaRESUMO
This paper consists of two parts on reversing-pulse electric birefringence (RPEB) signal patterns. The first is the theoretical formulation of two axially symmetric models coexisting in equilibrium in solution. The present RPEB theory is based on the original Tinoco-Yamaoka theory with classical electric dipole moments, which was recently modified and extended by Yamaoka, Sasai, and Kohno to include various electric and optical parameters and most importantly the ion-fluctuation dipole moment 0 or q<-1, the resultant patterns are often encountered with experimental signals. If -1
Assuntos
Eletricidade
, Modelos Teóricos
, Birrefringência
, Simulação por Computador
, Cinética
, Substâncias Macromoleculares
, Soluções/química
RESUMO
Steady-state and decay birefringence, expressed in terms of the optical phase retardation per cell length delta/d, was measured on beta-FeOOH in aqueous ionic media at 633 nm and at 25 degrees C by an electric square-pulse technique over a wide range of field strength E to ca. 6 kV/cm. The field-strength dependence of both delta/d and field-free rotational relaxation time tau was determined at the sample concentrations between 0.0011 and 0.055 g/L and in the 0.02-2.0 mM NaCl concentration range. Extrapolation of both delta/d and tau values to infinitely high fields (E(2)-->infinity) could yield birefringence- and weight-average quantities, respectively. Observed tau values were decreased at weak fields but leveled off to ca. 0.3 ms at very high fields due to a slight polydispersity regarding the length and volume of particles. The weight-average relaxation time tau(w) was calculated with Perrin's expressions theoretically from the length, width, and volume of beta-FeOOH particles estimated in the dried state from electron micrograph. These quantities were variously averaged. The size distribution was discussed in terms of observed discrete histogram and theoretical (Weibull and Lansing-Kraemer) distribution functions. The sign of observed delta/d value was always positive. The infinitely high-field (delta/d)(infinity) values and the reduced optical anisotropy factor Delta g/n were evaluated by fitting to theoretical orientation functions. The intrinsic birefringence (n3-n1) could be estimated with the mean refractive index n(p) reported in the literature. For the spindle-shaped particle with an axial ratio of ca. 4, the sign of Delta g/n is always positive, whereas the quantity (n3-n1) was either negative (n(p) > 2.35) or positive (n(p) < 2.05) in sign or nearly zero (ca. n(p) = 2.26), depending critically on the n(p) values.