Two dielectric relaxations, β_sp (1.5 MHz) and γ1_sp (7 MHz), have been detected on spectrin-based membrane skeleton (MS) of red blood cells (RBCs) using the plot of admittance changes at the spectrin denaturation temperature (Ivanov and Paarvanova in Bioelectrochemistry 110: 59–68, 2016, Electrochim Acta 317: 289–300, 2019a). In this study, we treated RBCs and RBC ghost membranes with agents that make membranes rigid and suppress membrane flicker, and studied the effect on β_sp and γ1_sp relaxations. Diamide (diazene dicarboxylic acid bis-(N,N-dimethylamide)) (up to 0.85 mM), taurine mustard (tris(2-chloroethyl)amine) (up to 2 mM), known to specifically cross-link and stiffen spectrin, and glutaraldehyde (up to 0.044%) all inhibited the relaxations in RBC ghost membranes. Similar inhibition was obtained resealing RBC ghost membranes with 2,3-diphosphoglicerate (up to 15 mM), binding WGA (wheat germ agglutinin) (up to 0.025 mg/ml) to exofacial aspect of RBCs, incubating RBCs in hypotonic (200 mOsm) and hypertonic (600–900 mOsm) media and depleting RBCs of ATP. By contrast, concanavalin A (1 mg/ml) and DIDS (4,4′-diiso-thiocyanato stilbene-2,2′-disulfonic acid) (75 μM, pH 8.2), both known to bind specifically band 3 integral protein of RBCs without effect on RBC membrane rigidity, did not affect the relaxations. We conclude there might be a relation between the strength of dielectric relaxations on MS spectrin and the deformability and flicker of RBC membrane.

使用血影蛋白变性温度下的导纳变化图，在基于血影蛋白的红细胞 (RBC) 膜骨架 (MS) 上检测到两种介电弛豫 β _sp (1.5 MHz) 和 γ1 _sp (7 MHz) (Ivanov)和 Paarvanova 在生物电化学 110: 59–68, 2016, Electrochim Acta 317: 289–300, 2019a)。在这项研究中，我们用使膜变硬并抑制膜闪烁的试剂处理红细胞和红细胞鬼膜，并研究了其对 β _sp和 γ1 _sp弛豫的影响。二酰胺（二氮烯二羧酸双-(N,N-二甲基酰胺)）（高达 0.85 mM）、牛磺酸芥末（三（2-氯乙基）胺）（高达 2 mM），已知可特异性交联和硬化血影蛋白，和戊二醛（高达0.044%）均抑制红细胞鬼膜的松弛。使用 2,3-二磷酸甘油酸（高达 15 mM）重新密封红细胞鬼膜，将 WGA（麦芽凝集素）（高达 0.025 mg/ml）结合到红细胞的外表面，在低渗（200 mOsm）下孵育红细胞，获得了类似的抑制作用和高渗 (600–900 mOsm) 介质并消耗红细胞的 ATP。相比之下，伴刀豆球蛋白 A (1 mg/ml) 和 DIDS（4,4'-二异硫氰酸二苯乙烯-2,2'-二磺酸）（75 μM，pH 8.2）均已知能特异性结合带 3 整合蛋白红细胞对红细胞膜的刚性没有影响，也不影响松弛。我们得出的结论是，MS 血影蛋白的介电弛豫强度与红细胞膜的变形性和闪烁之间可能存在关系。