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Sensing molecular organizational changes through the catalytic activity of acetylcholinesterase from erythrocyte membranes in Langmuir-Blodgett films.
Biochimica et Biophysica Acta (BBA) - Biomembranes ( IF 2.8 ) Pub Date : 2020-01-11 , DOI: 10.1016/j.bbamem.2020.183188 Iván Felsztyna 1 , Anahí V Turina 1 , María A Perillo 1 , Eduardo M Clop 1
Biochimica et Biophysica Acta (BBA) - Biomembranes ( IF 2.8 ) Pub Date : 2020-01-11 , DOI: 10.1016/j.bbamem.2020.183188 Iván Felsztyna 1 , Anahí V Turina 1 , María A Perillo 1 , Eduardo M Clop 1
Affiliation
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Langmuir films prepared from bovine erythrocyte membranes (LFBEM) were studied and transferred to alkylated glasses (Langmuir-Blodgett films, LBBEM) in order to assess the effects of membrane molecular packing on Bovine Erythrocyte Acetylcholinesterase (BEA) catalytic activity. Surface pressure (π) vs Area isotherms showed three 2D-transitions at ~7, ~18 and ~44 mN/m and a collapse pressure at πc = 49 mN/m. The 0-12-0 mN/m compression-decompression cycles resulted reversible while those 0-40-0 mN/m exhibited a significant hysteresis. Taken together, EFM, BAM and AFM images and the stability of the film after 3C-D cycles, we can suggest that over the air-water interface as well as over the silanized glass substrate the surface is mostly covered by a monolayer with a few particles dispersed. Acetylthiocholine hydrolysis was assayed with BEA in bovine erythrocyte membrane suspensions (SBEM) and in LBBEM packed at 10 (LBBEM,10) and 35 mN/m (LBBEM,35), which gave the following kinetic parameters: Vmax = 3.41 ± 0.15, 0.021 ± 0.002 and 0.030 ± 0.003 nmol.min-1·μg prot-1 and KM = 0.11 ± 0.02, 0.047 ± 0.017 and 0.026 ± 0.017 mM, respectively. Although from SBEM to LBBEM we lost active enzyme, the catalytic efficiency (Vmax/KM) increased ~750 times. Eugenol and 1,8-cineol inhibited BEA catalytic activity in LBBEM,35. Our results demonstrate the transmission of information between the membrane and the environment within the subphase immediately below the membrane, where anchored proteins are hosted. This was reflected by the membrane packing-induced modulation of BEA catalytic activity. Furthermore, LBBEM provides a proof of concept for the development of biosensors to screen new green pesticides acting through BEA interaction.
中文翻译：
通过Langmuir-Blodgett膜中红细胞膜的乙酰胆碱酯酶的催化活性来感知分子组织的变化。
研究了由牛红细胞膜（LFBEM）制备的Langmuir膜并将其转移至烷基化玻璃杯（Langmuir-Blodgett膜，LBBEM），以评估膜分子堆积对牛红细胞乙酰胆碱酯酶（BEA）催化活性的影响。表面压力（π）与面积等温线的关系在〜7，〜18和〜44 mN / m处显示了三个2D跃迁，在πc= 49 mN / m时出现了塌陷压力。0-12-0 mN / m压缩-减压循环可逆，而0-40-0 mN / m则显示出明显的滞后。综合考虑EFM，BAM和AFM图像以及3C-D循环后薄膜的稳定性，我们可以认为，在空气-水界面以及硅烷化玻璃基板上，表面大部分被单层覆盖，其中有一些颗粒分散。用BEA在牛红细胞膜悬浮液（SBEM）和以10（LBBEM，10）和35 mN / m（LBBEM，35）包装的LBBEM中测定乙酰硫胆碱水解的动力学参数：Vmax = 3.41±0.15，0.021 ±0.002和0.030±0.003 nmol.min-1·μgprot-1和KM分别为0.11±0.02、0.047±0.017和0.026±0.017 mM。尽管从SBEM到LBBEM，我们损失了活性酶，但催化效率（Vmax / KM）增加了约750倍。丁香酚和1,8-cineol抑制了LBBEM中的BEA催化活性[35]。我们的结果证明了膜与紧贴膜下面的亚相内环境之间的信息传递，其中固定了蛋白质。膜堆积诱导的BEA催化活性的调节反映了这一点。此外，
更新日期：2020-01-13
中文翻译：
通过Langmuir-Blodgett膜中红细胞膜的乙酰胆碱酯酶的催化活性来感知分子组织的变化。
研究了由牛红细胞膜（LFBEM）制备的Langmuir膜并将其转移至烷基化玻璃杯（Langmuir-Blodgett膜，LBBEM），以评估膜分子堆积对牛红细胞乙酰胆碱酯酶（BEA）催化活性的影响。表面压力（π）与面积等温线的关系在〜7，〜18和〜44 mN / m处显示了三个2D跃迁，在πc= 49 mN / m时出现了塌陷压力。0-12-0 mN / m压缩-减压循环可逆，而0-40-0 mN / m则显示出明显的滞后。综合考虑EFM，BAM和AFM图像以及3C-D循环后薄膜的稳定性，我们可以认为，在空气-水界面以及硅烷化玻璃基板上，表面大部分被单层覆盖，其中有一些颗粒分散。用BEA在牛红细胞膜悬浮液（SBEM）和以10（LBBEM，10）和35 mN / m（LBBEM，35）包装的LBBEM中测定乙酰硫胆碱水解的动力学参数：Vmax = 3.41±0.15，0.021 ±0.002和0.030±0.003 nmol.min-1·μgprot-1和KM分别为0.11±0.02、0.047±0.017和0.026±0.017 mM。尽管从SBEM到LBBEM，我们损失了活性酶，但催化效率（Vmax / KM）增加了约750倍。丁香酚和1,8-cineol抑制了LBBEM中的BEA催化活性[35]。我们的结果证明了膜与紧贴膜下面的亚相内环境之间的信息传递，其中固定了蛋白质。膜堆积诱导的BEA催化活性的调节反映了这一点。此外，
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