RESUMO
An understanding of the natural conformation of dolichol is important for the elucidation of the mechanism of protein glycosylation and dolichol's other as yet undisclosed biological functions. Since the molecular mechanics method has been shown to be well suited for the prediction of alcohol and alkene conformations, we have employed it to study the conformations of apparent least energy of dolichol-19 and smaller polymers of isoprene, namely, squalene, trans,trans-farnesol, and cis,cis-farnesol. Additionally, the small-angle X-ray scattering (SAXS) method was employed to determine the validity of the apparent least energy conformer of dolichol-19 derived by the molecular mechanics method. The results indicate that the solution conformation of dolichol-19 is comprised of a central coiled region flanked by two arms. The central coiled region has two and a half turns of dimensions 9.84 x 16.55 x 51.66 A3. The arms of dimensions 3.99 x 5.89 x 17.47 A3 and 4.49 x 9.23 x 11.14 A3 are approximately diametrically opposed. Measurement of the intrinsic viscosity of dolichol in both isopentyl alcohol and oleyl alcohol showed that the natural conformation of dolichol is capable of increasing solution fluidity (i.e., lowering solution viscosity). Thus, while examination of the conformation of dolichol in a membrane-mimetic solvent by SAXS is not possible, the quantitative measure of the effect of dolichol on solution viscosity (and thus solution fluidity) is possible. The results are consistent with dolichol acting as a membrane-fluidizing agent and provide the first quantitative measure of the effect of dolichol on solution fluidity of a membrane-mimetic solvent.
Assuntos
Dolicóis , Conformação Molecular , Estrutura Molecular , Espalhamento de Radiação , Estereoisomerismo , TermodinâmicaRESUMO
Solution characterization of heparin with high affinity (HA) and low affinity (LA) for antithrombin III was performed using the methods of small-angle x-ray scattering (SAXS), viscometry, and aqueous gel permeation chromatography (GPC). SAXS provided various topological parameters including the radius of gyration ([S2]1/2), radius of gyration of cross-section ([S2]q1/2), persistence length (a*), contour length (L), and mass parameters, e.g., overall molecular mass (Mr), and mass per unit length (Mq). The molecular weights of HA and LA pig mucosal heparins were found to be 14,900 and 11,500 and the respective radii of gyration were 40.1 and 33.6 A. The persistence lengths of HA and LA were 21.3 and 20.3 A, respectively. These parameters were compared to SAXS data of heparin [S. S. Stivala, M. Herbst, O. Kratky, and I. Pilz (1968) Arch. Biochem. Biophys. 127, 795-802] fractionated according to molecular weight only. It was found that the various experimental values of this heparin lie somewhere in between those of HA and LA heparins. It appears that there are no appreciable differences in the physico-chemical properties, including conformation, among the heparins in H2O at 25 degrees C.
Assuntos
Heparina/análise , Animais , Antitrombina III/metabolismo , Cromatografia em Gel , Heparina/metabolismo , Matemática , Peso Molecular , Ligação Proteica , Espalhamento de Radiação , Suínos , Viscosidade , Raios XAssuntos
Polissacarídeos Bacterianos , Luz , Espalhamento de Radiação , Streptococcus , Ultracentrifugação , ViscosidadeRESUMO
Levan produced by Streptococcus salivarius was fractionated into a series of 20 fractions of varying molecular weight. The range of intrinsic viscosities of the fractions was 0.07 to 0.18 dl/gm in water and 0.20 to 0.29 dl/gm in dimethyl sulfoxide. The molecular weight of the unfractionated leval determined by light scattering was 31.5 times 10-6. Small amounts of fatty acids and protein were found associated with levan.
