由在大肠杆菌中表达的枯草芽孢杆菌TetA(L)转运蛋白催化的四环素/H⁺反向转运和Na⁺/H⁺反向转运
Tetracycline/H+ antiport and Na+/H+ antiport catalyzed by the Bacillus subtilis TetA(L) transporter expressed in Escherichia coli.
作者信息
Guffanti A A, Krulwich T A
机构信息
Department of Biochemistry, Mount Sinai School of Medicine of CUNY, New York 10029, USA.
出版信息
J Bacteriol. 1995 Aug;177(15):4557-61. doi: 10.1128/jb.177.15.4557-4561.1995.
Abstract
The properties of TetA(L)-dependent tetracycline/proton and Na+/proton antiport were studied in energized everted vesicles of Escherichia coli transformed with a cloned tetA(L) gene (pJTA1) from Bacillus subtilis. Inhibition patterns by valinomycin and nigericin indicated that both antiports were electrogenic, in contrast to the tetracycline/proton antiport encoded by gram-negative plasmid tet genes. Tetracycline uptake in the everted system was dependent upon a divalent cation, with cobalt being the preferred one. The apparent Km for tetracycline was markedly increased at pH 8.5 versus pH 7.5, whereas the Vmax was unchanged. The much higher apparent Km for Na+ decreased at pH 8.5 relative to that at pH 7.5, as did the Vmax. Na+ did not affect tetracycline uptake, nor did Co2+ and/or tetracycline affect Na+ uptake; complex patterns of inhibition by amiloride and analogs thereof were observed.
摘要
利用克隆自枯草芽孢杆菌的tetA(L)基因（pJTA1）转化的大肠杆菌活力外翻囊泡，研究了依赖TetA(L)的四环素/质子和Na⁺/质子反向转运特性。缬氨霉素和尼日利亚菌素的抑制模式表明，与革兰氏阴性质粒tet基因编码的四环素/质子反向转运不同，这两种反向转运都是生电的。外翻系统中四环素的摄取依赖于二价阳离子，其中钴是首选。四环素的表观Km在pH 8.5时相对于pH 7.5显著增加，而Vmax不变。Na⁺的表观Km在pH 8.5时相对于pH 7.5显著降低，Vmax也降低。Na⁺不影响四环素摄取，Co²⁺和/或四环素也不影响Na⁺摄取；观察到阿米洛利及其类似物的复杂抑制模式。
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