Zhou Yong, Raphael Robert M
Department of Biochemistry and Cell Biology, Rice University, Houston, Texas, USA.
Biophys J. 2007 Apr 1;92(7):2451-62. doi: 10.1529/biophysj.106.096362. Epub 2006 Dec 15.
Solution pH affects numerous biological processes and some biological membranes are exposed to extreme pH environments. We utilized micropipette aspiration of giant unilamellar vesicles composed of 1-stearoyl-2-oleoyl-phosphatidylcholine to characterize the effect of solution pH (2-9) on membrane mechanical properties. The elastic area compressibility modulus was unaffected between pH 3 and 9 but was reduced by approximately 30% at pH 2. Fluorescence experiments utilizing the phase-sensitive probe Laurdan confirmed gel-phase characteristics at pH 2, explaining the reduction of membrane elasticity. The membrane bending stiffness, kc, increased by approximately 40% at pH 4 and pH 9 over the control value at pH 6.5. Electrophoretic mobility measurements indicate that these changes are qualitatively consistent with theoretical models that predict the effect of membrane surface charge density and Debye length on kc, substantiating a coupling between the mechanical and interfacial electrical properties of the membrane. The effect of pH on intramembrane electrical properties was examined by studying the spectral shifts of the potentiometric probe di-8 ANEPPS. The intramembrane (dipole) potential (Psid) increased linearly as the solution pH decreased in a manner consistent with the partitioning of hydroxide ions into the membrane. However, changes in Psid did not correlate with changes in kc. These mechanical and electrical studies lead to the conclusion that the effect of pH on membrane bending stiffness results from alterations in interfacial, as opposed to intramembrane, electrostatics.
溶液的pH值会影响众多生物过程,并且一些生物膜会暴露于极端pH环境中。我们利用微量移液器抽吸由1-硬脂酰-2-油酰磷脂酰胆碱组成的巨型单层囊泡,以表征溶液pH值(2 - 9)对膜力学性能的影响。弹性面积压缩模量在pH值为3至9之间不受影响,但在pH值为2时降低了约30%。利用相敏探针劳丹(Laurdan)进行的荧光实验证实了在pH值为2时的凝胶相特性,这解释了膜弹性的降低。膜弯曲刚度kc在pH值为4和pH值为9时比pH值为6.5时的对照值增加了约40%。电泳迁移率测量表明,这些变化在定性上与预测膜表面电荷密度和德拜长度对kc影响的理论模型一致,证实了膜的力学性质和界面电学性质之间的耦合。通过研究电位探针di - 8 ANEPPS的光谱位移来考察pH值对膜内电学性质的影响。膜内(偶极)电位(Psid)随着溶液pH值的降低呈线性增加,其方式与氢氧根离子在膜中的分配一致。然而,Psid的变化与kc的变化不相关。这些力学和电学研究得出结论,pH值对膜弯曲刚度的影响是由界面静电而非膜内静电的改变引起的。