Negoda Alexander, Kim Kwang-Jin, Crandall Edward D, Worden Robert M
Department of Chemical Engineering and Material Science, Michigan State University, East Lansing, MI 48824, USA.
Biochim Biophys Acta. 2013 Sep;1828(9):2215-22. doi: 10.1016/j.bbamem.2013.05.029. Epub 2013 Jun 5.
A diverse range of molecular interactions can occur between engineered nanomaterials (ENM) and biomembranes, some of which could lead to toxic outcomes following human exposure to ENM. In this study, we adapted electrophysiology methods to investigate the ability of 20nm polystyrene nanoparticles (PNP) to induce pores in model bilayer lipid membranes (BLM) that mimic biomembranes. PNP charge was varied using PNP decorated with either positive (amidine) groups or negative (carboxyl) groups, and BLM charge was varied using dioleoyl phospholipids having cationic (ethylphosphocholine), zwitterionic (phosphocholine), or anionic (phosphatidic acid) headgroups. Both positive and negative PNP induced BLM pores for all lipid compositions studied, as evidenced by current spikes and integral conductance. Stable PNP-induced pores exhibited ion selectivity, with the highest selectivity for K(+) (PK/PCl8.3) observed when both the PNP and lipids were negatively charged, and the highest selectivity for Cl(-) (PK/PCl0.2) observed when both the PNP and lipids were positively charged. This trend is consistent with the finding that selectivity for an ion in channel proteins is imparted by oppositely charged functional groups within the channel's filter region. The PK/PCl value was unaffected by the voltage-ramp method, the pore conductance, or the side of the BLM to which the PNP were applied. These results demonstrate for the first time that PNP can induce ion-selective pores in BLM, and that the degree of ion selectivity is influenced synergistically by the charges of both the lipid headgroups and functional groups on the PNP.
工程纳米材料(ENM)与生物膜之间可能发生多种分子相互作用,其中一些相互作用在人类接触ENM后可能导致毒性后果。在本研究中,我们采用电生理学方法来研究20纳米聚苯乙烯纳米颗粒(PNP)在模拟生物膜的模型双层脂质膜(BLM)中诱导形成孔的能力。通过用带正电(脒基)基团或带负电(羧基)基团修饰PNP来改变PNP的电荷,并用具有阳离子(乙基磷脂酰胆碱)、两性离子(磷脂酰胆碱)或阴离子(磷脂酸)头部基团的二油酰磷脂来改变BLM的电荷。对于所有研究的脂质组成,带正电和带负电的PNP均能诱导BLM形成孔,电流尖峰和积分电导率证明了这一点。稳定的PNP诱导孔表现出离子选择性,当PNP和脂质均带负电时,观察到对K(+)的选择性最高(PK/PCl8.3),而当PNP和脂质均带正电时,观察到对Cl(-)的选择性最高(PK/PCl0.2)。这一趋势与以下发现一致,即通道蛋白中离子的选择性是由通道过滤区域内带相反电荷的官能团赋予的。PK/PCl值不受电压斜坡法、孔电导率或施加PNP的BLM一侧的影响。这些结果首次证明PNP可在BLM中诱导形成离子选择性孔,并且离子选择性程度受到脂质头部基团和PNP上官能团电荷的协同影响。