Ballas S K, Mohandas N, Marton L J, Shohet S B
Proc Natl Acad Sci U S A. 1983 Apr;80(7):1942-6. doi: 10.1073/pnas.80.7.1942.
Using a laser diffraction technique, we have studied the effects of putrescine, spermidine, and spermine, the three physiologic polyamines, on the deformability and mechanical stability of human erythrocyte membranes. Ghosts resealed with polyamines were subjected to high fluid shear stress in an ektacytometer. All polyamines increased the membrane shear modulus (decreased deformability) in a concentration- and time-dependent manner. The order of effectiveness was spermine greater than spermidine greater than putrescine. At 10 microM, spermine appreciably decreased membrane deformability. For the measurement of membrane mechanical stability, resealed ghosts were subjected to constant high shear stress in the ektacytometer and deformability was continuously recorded as the deformable ghosts fragmented into rigid spherical vesicles. Polyamines, especially spermine, caused a noticeable increase in the t1/2 for fragmentation. These effects could not be ascribed to proteolysis or Ca2+-induced transglutamination. That the effects of polyamines were specific and not simply due to their positive charge was demonstrated by the finding that Ca2+ and Mg2+ destabilized the erythrocyte membrane as evidenced by decreasing the t1/2 for fragmentation. Extracellular polyamines were not effective except under conditions that caused significant accumulation inside the cell. The data indicate that intracellular physiologic polyamines, especially spermine, decrease erythrocyte membrane deformability and stabilize the membrane skeleton, making it more resistant to fragmentation.
我们运用激光衍射技术,研究了三种生理性多胺——腐胺、亚精胺和精胺对人红细胞膜变形性和机械稳定性的影响。用多胺重新封闭的血影细胞在激光衍射血细胞仪中承受高流体剪切应力。所有多胺均以浓度和时间依赖性方式增加膜剪切模量(降低变形性)。有效性顺序为精胺>亚精胺>腐胺。在10微摩尔浓度时,精胺显著降低膜变形性。为测量膜的机械稳定性,将重新封闭的血影细胞在激光衍射血细胞仪中承受恒定的高剪切应力,并在可变形的血影细胞破碎成刚性球形囊泡时持续记录变形性。多胺,尤其是精胺,使破碎的t1/2显著增加。这些效应不能归因于蛋白水解或Ca2+诱导的转谷氨酰胺化。Ca2+和Mg2+使红细胞膜不稳定,表现为降低破碎的t1/2,这一发现证明多胺的作用具有特异性,并非仅仅因其正电荷所致。除了在导致细胞内大量蓄积的条件下,细胞外多胺无效。数据表明,细胞内生理性多胺,尤其是精胺,会降低红细胞膜变形性并稳定膜骨架,使其更抗破碎。