Shih Y L, Wang W P, Zhang H, Wang K Y, Guo M
Shanghai Institute of Physiology, Academia Sinica.
Sheng Li Xue Bao. 1992 Apr;44(2):142-8.
Pinellia ternata lectin (PTL), a protein exhibiting hemagglutination activity and carbohydrate binding specificity to mannan was purified from rhizome of Pinellia ternata. In this work the actions of PTL on artificial lipid bilayer were investigated by means of the two-compartment system of Mueller and Rudin. The lipid bilayer with resistance more than 10G omega was formed by a solution of lecithin and cholesterol (20 mg/ml and 5 mg/ml respectively) in N-decane. The electrical properties of the lipid bilayer were investigated in voltage clamp mode. Several minutes after the addition of PTL (2 micrograms/ml) in one compartment the channel-like noise as well as a decrease of the resistance of the bilayer were observed. These actions were inhibited by mannan significantly. The resistance increase of the bilayer with PTL-channels could be observed from 2G omega to control level (greater than or equal to 10 G omega) immediately after addition of 40 micrograms/ml mannan. The discrete conduction steps were recorded at low concentration of PTL and at low holding potential. The predominant unit conductance was 35pS in symmetric KCl solution of 100 mmol/L. The selectivity of PTL-channel was estimated from Goldman-Hodgkin-Katz equation by measurement of the reversal potential in an asymmetrical salt solution. The results showed that PTL-channels were cation selective.
半夏凝集素(PTL)是一种从半夏根茎中纯化得到的具有血凝活性且对甘露聚糖具有碳水化合物结合特异性的蛋白质。在本研究中,采用穆勒和鲁丁的双室系统研究了PTL对人工脂质双层的作用。用卵磷脂和胆固醇(分别为20mg/ml和5mg/ml)在正癸烷中的溶液形成电阻大于10GΩ的脂质双层。在电压钳模式下研究脂质双层的电学性质。在一个隔室中加入PTL(2μg/ml)几分钟后,观察到类似通道的噪声以及双层电阻的降低。这些作用被甘露聚糖显著抑制。加入40μg/ml甘露聚糖后,立即观察到带有PTL通道的双层电阻从2GΩ增加到对照水平(大于或等于10GΩ)。在低浓度的PTL和低保持电位下记录到离散的传导步骤。在100mmol/L的对称KCl溶液中,主要单位电导为35pS。通过在不对称盐溶液中测量反转电位,根据戈德曼-霍奇金- Katz方程估算PTL通道的选择性。结果表明,PTL通道具有阳离子选择性。