Kupisz Kamila, Trebacz Kazimierz, Gruszecki Wiesław I
Department of Biophysics, Institute of Biology and Biochemistry, Maria Curie-Skłodowska University, Lublin, 20-033, Poland.
Department of Biophysics, Institute of Physics, Maria Curie-Skłodowska University, Lublin, 20-031, Poland.
Physiol Plant. 2015 Jul;154(3):349-57. doi: 10.1111/ppl.12288. Epub 2014 Nov 3.
Our previous study has shown that the liverwort Conocephalum conicum generates action potentials (APs) in response to both temperature drop and menthol, which are also activators of the TRPM8 (transient receptor potential melastatin 8) receptor in animals. Not only similarities but also differences between electrical reactions to menthol and cooling observed in the liverwort aroused our interest in the action of menthol at the molecular level. Patch-clamp investigations have shown that menthol causes a reduction of current flowing through slow vacuolar (SV) channels to 29 ± 10% of the initial value (n = 9); simultaneously, it does not influence magnitudes of currents passing through a single SV channel. This may point to an unspecific interaction between menthol and the lipid phase of the membrane. An influence of menthol on lipid organization in membranes was investigated in two-component monomolecular layers formed with menthol and dipalmitoylphosphatidylcholine (DPPC) at the argon-water interface. Analyses of the mean molecular area parameters vs the molar fraction of the menthol component have shown over-additivity (approximately 20 Å(2) ) in the region of high molar fractions of menthol. Infrared absorption spectroscopy studies have shown that menthol, most probably, induces breaking of a hydrogen bond network formed by ester carbonyl groups and water bridges in the lipid membrane and binds to the polar head group region of DPPC. We conclude that the disruption in the lipid phase of the membrane influences ion channels and/or pumps and subsequently causes generation of APs in excitable plants such as C. conicum.
我们之前的研究表明,地钱(Conocephalum conicum)会响应温度下降和薄荷醇产生动作电位(APs),而温度下降和薄荷醇在动物体内也是瞬时受体电位香草酸亚型8(TRPM8)受体的激活剂。在地钱中观察到的对薄荷醇和冷却的电反应之间,不仅存在相似性,也存在差异,这激发了我们在分子水平上对薄荷醇作用的兴趣。膜片钳研究表明,薄荷醇会使通过慢液泡(SV)通道的电流降低至初始值的29±10%(n = 9);同时,它不会影响通过单个SV通道的电流大小。这可能表明薄荷醇与膜的脂质相之间存在非特异性相互作用。在氩 - 水界面上由薄荷醇和二棕榈酰磷脂酰胆碱(DPPC)形成的双组分单分子层中,研究了薄荷醇对膜中脂质组织的影响。对平均分子面积参数与薄荷醇组分摩尔分数的分析表明,在高摩尔分数的薄荷醇区域存在超加和性(约20 Ų)。红外吸收光谱研究表明,薄荷醇很可能会诱导脂质膜中由酯羰基和水桥形成的氢键网络断裂,并与DPPC的极性头部基团区域结合。我们得出结论,膜脂质相的破坏会影响离子通道和/或泵,随后导致像地钱这样的可兴奋植物产生动作电位。