The Low-Resolution Structure of Nascent High Density Lipoprotein Reconstituted with DMPC With and Without Cholesterol Reveals A Mechanism for Particle Expansion

High density lipoproteins (HDL) are athero-protective particles under investigation as potential therapeutic agents for cardiovascular disease. We applied small angle neutron scattering (SANS) with contrast variation to obtain the low resolution structure of nascent HDL (nHDL) reconstituted with dimyristoyl phosphatidyl choline (DMPC), apoA1:DMPC (1:80, mol:mol). The overall shape of the entire particle is discoidal, with low resolution architecture of apoA1 visualized as an open, contorted, and slightly out of plane structure with three arms, while the low resolution shape of the lipid phase is an oblate ellipsoid that fits well within the protein shape. Modeling studies incorporating the SANS data indicate that apoA1 within the lipoprotein is folded onto itself, making a hairpin, which was also confirmed independently by both cross-linking mass spectrometry and hydrogen-deuterium exchange mass spectrometry analyses. The open conformation of apoA1 observed coupled with the lipid shape indicate that the lipid is predominantly a bilayer with a small micelle domain between the open apoA1 arms. Collectively, these studies demonstrate that full length apoA1 retains an open architecture that is dictated by its lipid cargo. This configuration may help accommodate potential changing lipid cargo content of the particle by quantized expansion of hairpin structures in apoA

An Abundant Dysfunctional Apolipoprotein A1 in Human Atheroma

Recent studies have indicated that high-density lipoproteins (HDLs) and their major structural protein, apolipoprotein A1 (apoA1), recovered from human atheroma are dysfunctional and are extensively oxidized by myeloperoxidase (MPO). In vitro oxidation of either apoA1 or HDL particles by MPO impairs their cholesterol acceptor function. Here, using phage display affinity maturation, we developed a high-affinity monoclonal antibody that specifically recognizes both apoA1 and HDL that have been modified by the MPO-H2O2-Cl− system. An oxindolyl alanine (2-OH-Trp) moiety at Trp72 of apoA1 is the immunogenic epitope. Mutagenesis studies confirmed a critical role for apoA1 Trp72 in MPO-mediated inhibition of the ATP-binding cassette transporter A1 (ABCA1)-dependent cholesterol acceptor activity of apoA1 in vitro and in vivo. ApoA1 containing a 2-OH-Trp72 group (oxTrp72-apoA1) is in low abundance within the circulation but accounts for 20% of the apoA1 in atherosclerosis-laden arteries. OxTrp72-apoA1 recovered from human atheroma or plasma is lipid poor, virtually devoid of cholesterol acceptor activity and demonstrated both a potent proinflammatory activity on endothelial cells and an impaired HDL biogenesis activity in vivo. Elevated oxTrp72-apoA1 levels in subjects presenting to a cardiology clinic (n = 627) were associated with increased cardiovascular disease risk. Circulating oxTrp72-apoA1 levels may serve as a way to monitor a proatherogenic process in the artery wall

Deeper understanding of non-linear geodetic data inversion using a quantitative sensitive analysis

A quantitative global sensitivity analysis (SA) is applied to the non-linear inversion of gravity changes and displacement data which measured in an active volcanic area. The common inversion of this data is based on the solution of the generalized Navier equations which couples both types of observation, gravity and displacement, in a homogeneous half space. The sensitivity analysis has been carried out using Sobol’s variance-based approach which produces the total sensitivity indices (TSI), so that all interactions between the unknown input parameters are taken into account. Results of the SA show quite different sensitivities for the measured changes as they relate to the unknown parameters for the east, north and height component, as well as the pressure, radial and mass component of an elastic-gravitational source. The TSIs are implemented into the inversion in order to stabilize the computation of the unknown parameters, which showed wide dispersion ranges in earlier optimization approaches. Samples which were computed using a genetic algorithm (GA) optimization are compared to samples in which the results of the global sensitivity analysis are integrated by a reweighting of the cofactor matrix in the objective function. The comparison shows that the implementation of the TSI’s can decrease the dispersion rate of unknown input parameters, producing a great improvement the reliable determination of the unknown parameters.Peer reviewe