Noninvasive neutron scattering measurements reveal slower cholesterol transport in model lipid membranes.

Proper cholesterol transport is essential to healthy cellular activity and any abnormality can lead to several fatal diseases. However, complete understandings of cholesterol homeostasis in the cell remains elusive, partly due to the wide variability in reported values for intra- and intermembrane cholesterol transport rates. Here, we used time-resolved small-angle neutron scattering to measure cholesterol intermembrane exchange and intramembrane flipping rates, in situ, without recourse to any external fields or compounds. We found significantly slower transport kinetics than reported by previous studies, particularly for intramembrane flipping where our measured rates are several orders of magnitude slower. We unambiguously demonstrate that the presence of chemical tags and extraneous compounds employed in traditional kinetic measurements dramatically affect the system thermodynamics, accelerating cholesterol transport rates by an order of magnitude. To our knowledge, this work provides new insights into cholesterol transport process disorders, and challenges many of the underlying assumptions used in most cholesterol transport studies to date.

Progress in the development of phase-sensitive neutron reflectometry methods.

It has been a number of years since phase-sensitive specular neutron reflectometry (PSNR) methods employing reference layers were first introduced to help remove the ambiguity inherent in the reconstruction of scattering length density (SLD) depth profiles (Majkrzak, C. F.; Berk, N. F. Physica B 2003, 336, 27) from specular reflectivity measurements. Although a number of scientific applications of PSNR techniques have now been successfully realized (Majkrzak, C. F.; Berk, N. F.; Perez-Salas, U. A. Langmuir 2003, 19, 7796 and references therein), in certain cases practical difficulties remain. In this article, we describe possible solutions to two specific problems: (1) the need for explicit, detailed knowledge of the SLD profile of a given reference layer of finite thickness; and (2) for a reference layer of finite thickness in which only two density variations are possible, how to identify which of two mathematical solutions corresponds to the true physical structure.

Persistence length changes dramatically as RNA folds.

We determine the persistence length l(p) for a bacterial group I ribozyme as a function of concentration of monovalent and divalent cations by fitting the distance distribution functions P(r) obtained from small angle x-ray scattering intensity data to the asymptotic form of the calculated P(WLC)(r) for a wormlike chain. The l(p) values change dramatically over a narrow range of Mg(2+) concentration from approximately 21 Angstroms in the unfolded state (U) to approximately 10 Angstroms in the compact (I(C)) and native states. Variations in l(p) with increasing Na(+) concentration are more gradual. In accord with the predictions of polyelectrolyte theory we find l(p) alpha 1/kappa(2) where kappa is the inverse Debye-screening length.