RESUMO
Cholesterol constitutes â¼30-40% of the mammalian plasma membrane, a larger fraction than of any other single component. It is a major player in numerous signaling processes as well as in shaping molecular membrane architecture. However, our knowledge of the dynamics of cholesterol in the plasma membrane is limited, restricting our understanding of the mechanisms regulating its involvement in cell signaling. Here, we applied advanced fluorescence imaging and spectroscopy approaches on in vitro (model membranes) and in vivo (live cells and embryos) membranes as well as in silico analysis to systematically study the nanoscale dynamics of cholesterol in biological membranes. Our results indicate that cholesterol diffuses faster than phospholipids in live membranes, but not in model membranes. Interestingly, a detailed statistical diffusion analysis suggested two-component diffusion for cholesterol in the plasma membrane of live cells. One of these components was similar to a freely diffusing phospholipid analogue, whereas the other one was significantly faster. When a cholesterol analogue was localized to the outer leaflet only, the fast diffusion of cholesterol disappeared, and it diffused similarly to phospholipids. Overall, our results suggest that cholesterol diffusion in the cell membrane is heterogeneous and that this diffusional heterogeneity is due to cholesterol's nanoscale interactions and localization in the membrane.
Assuntos
Membrana Celular/química , Colesterol/análise , Simulação de Dinâmica Molecular , Nanotecnologia , Animais , Células CHO , Membrana Celular/metabolismo , Células Cultivadas , Colesterol/metabolismo , Cricetulus , Difusão , Feminino , Masculino , Método de Monte Carlo , Espectrometria de Fluorescência , Peixe-ZebraRESUMO
Coarse-grained (CG) modeling has emerged as a promising tool to bridge the gap between the temporal and spatial scales of all-atom (AA) simulations and those of many important biological processes. Resolution exchange, a variant of the replica exchange method, combines the efficiency of CG simulation and the accuracy of AA simulation by swapping configurations between AA and CG simulations. The crucial step in a resolution exchange move is to rigorously reconstruct the high-resolution system from models at coarser resolutions. In this paper, configurational-bias Monte Carlo is adopted as a general method to rebuild the missing degrees of freedom rigorously for CG models and for the first time combined with resolution exchange. The new approach is demonstrated on an alkane and a peptide system. It is found that the efficiency of resolution exchange depends significantly on the quality of the CG model.