Roy Sudipa Saha, Sen Gargi, Biswas Tuli
Indian Institute of Chemical Biology, 4 Raja S. C. Mullick Road, Kolkata 700 032, India.
Arch Biochem Biophys. 2005 Apr 1;436(1):121-7. doi: 10.1016/j.abb.2005.01.015.
Membrane destabilization in erythrocytes plays an important role in the premature hemolysis and development of anemia during visceral leishmaniasis (VL). Marked degradation of the anion channel protein band 3 is likely to allow modulation of anion flux across the red cell membrane in infected animals. The present study describes the effect of structural modification of band 3 on phosphate transport in VL using (31)P NMR. The result showed progressive decrease in the rate and extent of phosphate transport during the post-infection period. Interdependence between the intracellular ionic levels seems to be a determining factor in the regulation of anion transport across the erythrocyte membrane in control and infected conditions. Infection-induced alteration in band 3 made the active sites of transport more susceptible to binding with amino reactive agents. Inhibition of transport by oxidation of band 3 and subsequent reversal by reduction using dithiothreitol suggests the contribution of sulfhydryl group in the regulation of anion exchange across the membrane. Quantitation of sulfhydryl groups in the anion channel protein showed the inhibition to be closely related to the decrease of sulfhydryl groups in the infected hamsters. Downregulation of phosphate transport during leishmanial infection may be ascribed to the sulfhydryl modification of band 3 resulting in the impaired functioning of this protein under the diseased condition.
红细胞膜去稳定化在内脏利什曼病(VL)期间的过早溶血和贫血发展中起重要作用。阴离子通道蛋白带3的显著降解可能会调节受感染动物红细胞膜上的阴离子通量。本研究利用(31)P核磁共振描述了带3结构修饰对VL中磷酸盐转运的影响。结果显示,感染后期间磷酸盐转运的速率和程度逐渐降低。在对照和感染条件下,细胞内离子水平之间的相互依赖性似乎是调节红细胞膜阴离子转运的一个决定性因素。感染诱导的带3改变使转运的活性位点更容易与氨基反应剂结合。通过氧化带3抑制转运,随后使用二硫苏糖醇还原使其逆转,这表明巯基在调节跨膜阴离子交换中起作用。对阴离子通道蛋白中巯基的定量显示,抑制作用与感染仓鼠中巯基的减少密切相关。利什曼原虫感染期间磷酸盐转运的下调可能归因于带3的巯基修饰,导致该蛋白在患病条件下功能受损。