N-hydroxysulfosuccinimido active esters and the L-(+)-lactate transport protein in rabbit erythrocytes.
N-hydroxysulfosuccinimido active esters and the L-(+)-lactate transport protein in rabbit erythrocytes.
复制标题
兔红细胞中的 N-羟基磺基琥珀酰亚胺活性酯和 L-( )-乳酸转运蛋白。
DOI：
10.1021/bi00355a012
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发表时间：
1986
期刊：
影响因子：
2.9
通讯作者：
Jennings,ML
中科院分区：
文献类型：
--
作者：
Donovan,JA;Jennings,ML
爱荷华州爱荷华市爱荷华大学生理和生物物理系，52242收到1985年8月27日摘要：W-羟基磺基琥珀酰亚胺酯强烈抑制兔红细胞L-(+)-乳酸转运蛋白，可能是通过酰化转运蛋白上的氨基来实现的。使用双(磺基琥珀酰亚胺)琥珀酸酯(BS3)、双(磺基琥珀酰亚胺)己二酸(BS2A)、双(磺基琥珀酰亚胺)二硫代丙酸酯(丙酸酯)和各种单羧酸酯进行的乳酸转运研究表明，乳酸转运蛋白的表面氨基是乳酸转运所必需的。此外，还原甲基化研究表明，即使在修饰的氨基中保留正电荷，转运也会受到强烈的抑制。在pH和lt；6时，带3介导的无机阴离子转运在BS3处理的细胞中被促进，而在pH和gt；6时，它被抑制。BSM对L-(+)-乳酸转运的抑制作用不具有这种pH依赖性。BS3减少了氚4，4‘-二异硫氰酸盐-2，2-二氢二苯乙烯二磺酸([~3H]H_2DID)和氚双(磺基琥珀酰亚胺)己二酸([~H]BS2A)对40-50 kDa膜多肽(带R)的标记。氚琥珀酸亚胺(S2[~3H]乙酸酯)也标记R带，在一定的浓度范围内，S2乙酸酯以剂量依赖的方式抑制乳酸的转运。BS3是一种已知的非本意蛋白质交联剂。S_2醋酸酯以一种H_2DDS抑制机制渗透到兔红细胞膜。BS3是胞外胰凝乳酶产生的兔带3蛋白水解物的交联物。这些标记实验支持带R和特定的单羧酸转运之间的联系。
Department of Physiology and Biophysics, The University of Iowa, Iowa City, Iowa 52242 Received August 27, 1985 abstract: Esters of W-hydroxysulfosuccinimide strongly inhibit L-(+)-lactate transportin rabbit erythrocytes, probably by acylating amino groups on the transport protein. Lactate transport studies using bis (sulfosuccinimido) suberate (BS3), bis (sulfosuccinimido) adipate (BS2A), bis (sulfosuccinimido) dithiobis (pro-pionate), and a variety of monocarboxylate esters suggest that an exofacial amino group of the lactate transport protein is essential for lactate transport. Also, reductive methylation studies show that even when positive charge is preserved in modified amino groups, the transport is strongly inhibited. At pH< 6, band 3 mediated inorganic anion transport is enhanced in BS3-treated cells, while at pH> 6, it is inhibited. BSMnduced inhibition of L-(+)-lactate transport does not have thispH dependence. BS3 reduces the labeling of a 40-50-kDa membrane polypeptide (band R) by tritiated 4, 4'-diisothiocyanato-2, 2-dihydrostilbenedisulfonate ([3H] H2DIDS) and by tritiated bis (sulfosuccinimido) adipate ([3H] BS2A). Tritiated sulfosuccinimido acetate (S2 [3H] acetate) also labels band R, over a range of concentrations where lactate transport is inhibited in a dose-dependent manner by S2acetate. BS3 is a known impermeant protein cross-linker. S2acetate permeates rabbit red cell membranes by an H2DIDS-inhibitable mechanism. BS3 cross-links the proteolytic fragments of rabbit band 3 produced by extracellular chymotrypsin. These labeling experiments support an association between band R and specific monocarboxylate transport.