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α1-酸性糖蛋白依赖性水和离子在棕榈酰-L-油酰磷脂酰胆碱脂质体中渗透转运的动力学及作用

Kinetics and role of alpha 1-acid glycoprotein-dependent osmotic transport of water and ions in palmitoyl-L-oleoyl phosphatidylcholine liposomes.

作者信息

Neitchev V, Kostova E, Goldenberg M, Doumanova L

机构信息

Institute of Biophysics, Bulgarian Academy of Sciences, Sofia, Bulgaria.

出版信息

Int J Biochem Cell Biol. 1997 Apr;29(4):689-701. doi: 10.1016/s1357-2725(96)00157-4.

Abstract

alpha 1-Acid glycoprotein isolated from human blood plasma is known to influence cell permeability, although the mechanisms of this process are unclear. Here, the glycoprotein effects on the permeability of osmotically stressed phospholipid liposomes are studied as a model of membrane permeability. Liposomes containing glycoprotein were found to be osmotically sensitive to water and chloride salts of some monovalent (Na+, K+) and bivalent (Mg2+, Ca2+) ions. The permeations of these substances were determined by light-scattering measurements of the volume changes in liposomes after mixing with hyperosmotic solutions of chloride salts. The time courses of scattered light were recorded by means of stopped-flow spectrophotometry. Two processes were studied: the fast water outflow from liposomes and slower ion permeations through the lipid membrane. The second order permeation rate constants were determined at different glycoprotein concentrations for both processes. Values from 66 to 250 x 10(3) for water outflow and 2-500 M-1 sec-1 for the different ion permeations were obtained in order to characterize the permeations of solutes across the lipid membrane. The apparent activation energies also were calculated between 18 and 33 degrees C. The mercurial sulphydryl reagent pCMBS inhibited the ion permeations in the slow phase. When pCMBS was present in this phase, higher activation energies were obtained, indicating more difficult permeations. An interpretation of these results is that membrane permeability is mediated by aqueous pores. Membrane selectivity to monovalent metal ions also was demonstrated, but no correlation was observed between the ion radius of the corresponding metal cation and permeation rate constants. The discovery of non-specific pores in liposomes containing glycoprotein shows that they can serve as vehicles for the water and ions in the processes of passive transport through lipid membranes.

摘要

从人血浆中分离出的α1-酸性糖蛋白已知会影响细胞通透性,尽管这一过程的机制尚不清楚。在此,研究了该糖蛋白对渗透应激磷脂脂质体通透性的影响,以此作为膜通透性的模型。发现含有糖蛋白的脂质体对水以及某些单价(Na +、K +)和二价(Mg2 +、Ca2 +)离子的氯化物盐具有渗透敏感性。这些物质的渗透通过与氯化物盐的高渗溶液混合后脂质体体积变化的光散射测量来确定。散射光的时间进程通过停流分光光度法记录。研究了两个过程:脂质体中快速的水流流出和较慢的离子通过脂质膜的渗透。针对这两个过程,在不同糖蛋白浓度下测定了二级渗透速率常数。得到水流出的速率常数在66至250×10³之间,不同离子渗透的速率常数在2至500 M⁻¹ s⁻¹之间,以便表征溶质跨脂质膜的渗透。还计算了18至33摄氏度之间的表观活化能。汞巯基试剂对氯汞苯甲酸(pCMBS)在慢相中抑制离子渗透。当在此相中存在pCMBS时,获得了更高的活化能,表明渗透更困难。对这些结果的一种解释是膜通透性由水通道介导。还证明了膜对单价金属离子的选择性,但未观察到相应金属阳离子的离子半径与渗透速率常数之间的相关性。在含有糖蛋白的脂质体中发现非特异性孔表明它们可作为脂质膜被动运输过程中水和离子的载体。

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