Water permeability of the chromaffin granule membrane

NMR spin-lattice relaxation rates of solvent protons have been used to measure the water permeability coefficient of the chromaffin granule membrane. The technique involves labeling the chromaffin granule interior with Mn+2, which provides an efficient relaxation pathway for intravesicular solvent protons. Added Mn+2 spontaneously accumulates in the chromaffin granule matrix in the presence of the divalent cation-specific ionophore A23187 and is maintained against a large concentration gradient. In this way, the internal proton relaxation rate is readily augmented to values some 10(2)-10(3) times greater than that in the extravesicular water space. Transmembranal water transport permits solvent protons in the extravesicular water space, in which most of the observed NMR signal orginates, to sample the highly relaxive environment of the chromaffin granule matrix. By this process, water permeation shortens the observed relaxation rate. The diffusive water permeability coefficient of the chromaffin granule membrane has been measured over the temperature range 0–38 degrees C. The permeability coefficient measured at 25 degrees C is comparable to a previously reported value for planar lipid bilayers composed of ox brain lipids and cholesterol (Pd approximately equal to 0.37–0.53 10(-3)) cm X s-1 at 25 degrees C) but is substantially less than values for the plasma membranes of erythrocytes and Chlorella. Hypothesized hydrophilic "pores," thought to provide parallel permeation pathways in the latter membranes, appear to be absent in chromaffin granule membranes. The water permeation rate exhibits Arrhenius temperature behavior and does not reflect a phase transition at 32 degrees-34 degrees C observed previously in ESR spin-label studies of chromaffin granule ghosts