Water Diffusion in Bicelles and the Mixed Bicelle Model

To test a prediction of the mixed bicelle model, stimulated echo (STE) pulsed field gradient (PFG) 1H nuclear magnetic resonance (NMR) measurements of water diffusion between and across bicellar lamellae were performed in positively and negatively magnetically aligned bicelles, composed of mixtures of DHPC (1,2-dihexanoyl-sn-glycero-3-phosphocholine) and DMPC (1,2-dimyristoyl-sn-glycero-3-phosphocholine), as a function of temperature and of the proportion of added short-chain lipid DHPC. 31P NMR spectra obtained for each situation confirmed that the DHPC undergoes fast exchange between curved and planar regions as per the mixed bicelle model and permitted an estimate of the proportion of the two DHPC populations. Water diffusion across the bicellar lamellae was shown to scale directly with q*, the fraction of edge versus planar phospholipid, rather than simply the ratio q, the global fraction of long-chain to short-chain phospholipid. Geometric modeling of the dependence of water diffusion on q* suggested an upper limit of 400 Å for the size of DHPC-rich toroidal perforations within the bicelle lamellae. These findings constitute an independent confirmation of the mixed bicelle model in which DHPC is not confined to edge regions but enjoys, instead, a finite miscibility with DMPC.