PERMEABILITY CHARACTERISTICS OF ERYTHROCYTE-GHOSTS PREPARED UNDER ISOIONIC CONDITIONS BY A GLYCOL-INDUCED OSMOTIC LYSIS
PERMEABILITY CHARACTERISTICS OF ERYTHROCYTE-GHOSTS PREPARED UNDER ISOIONIC CONDITIONS BY A GLYCOL-INDUCED OSMOTIC LYSIS
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DOI：
10.1016/0005-2736(77)90269-3
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发表时间：
1977-01-01
期刊：
影响因子：
--
通讯作者：
MICHELL, RH
中科院分区：
文献类型：
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作者：
BILLAH, MM;FINEAN, JB;MICHELL, RH
在清除所有Hb所需的5-7次裂解后，对大分子（人红细胞中）如葡聚糖（平均MW 70，000）的不渗透性立即自发恢复。对较小分子如MgATP 2-、[3 H]肌醇和[14 C]胆碱的渗透性最初很高，但在37 ℃温育后可大大降低。C下保温1h。这种重新密封的程度随着血影所经受的溶解次数的增加而减少。pH、Ca ~（2+）浓度和二价阳离子螯合剂对Hb的去除率和对小分子物质的渗透性均有显著影响。在pH 7.0（0 ℃）的碱性离子介质（130 mM KCl，10 mM NaCl，2 mM MgCl 2，10 mM N-2-羟乙基哌嗪-N ″-2-乙磺酸（HEPES））中制备的血影中实现最大的再密封。C）和Ca水平约10-5 M。酸性pH促进Hb的去除，而二价阳离子螯合剂的存在减缓了其释放。在第一次裂解期间，通过装载有K+的血影保留K+，随后在37 ℃温育。C是实质性的，但很少的K+可以保留在无Hb鬼。渗透性的鬼1裂解后，K+的温度，pH值，Ca 2+浓度和二价阳离子螯合剂的存在下的影响。
Impermeability to large molecules [in human erythrocytes] such as dextran (average MW 70,000) was restored immediately and spontaneously after each of the 5-7 lyses that were required to remove all of the Hb. Permeabilities to smaller molecules such as MgATP2-, [3H]inositol and [14C]choline were initially high but could be greatly reduced by incubation at 37.degree. C for 1 h. The extent of such resealing decreased as the number of lyses to which the ghosts were subjected increased. Both removal of Hb and permeabilities to small molecules were affected significantly by pH, Ca2+ concentrations and divalent cation chelators. Maximum resealing was achieved in ghosts prepared in the basic ionic medium (130 mM KCl, 10 mM NaCl, 2 mM MgCl2, 10 mM N-2-hydroxyethylpiperazine-N''-2-ethanesulfonic acid (HEPES)) at pH 7.0 (0.degree. C) and with a Ca level around 10-5 M. Acidic pH facilitated the removal of Hb, while the presence of divalent cation chelators slowed down its release. Retention of K+ by ghosts loaded with K+ during the 1st lysis and subsequently incubated at 37.degree. C was substantial but little K+ could be retained within the Hb-free ghosts. Permeability of the ghosts to K+ after 1 lysis was affected by temperature, pH, Ca2+ concentrations and by the presence of divalent cation chelators.