适应淡水和海水的鳗鱼（欧洲鳗鲡）鳃中诱导型钠钾离子依赖性三磷酸腺苷酶的性质和特性。

The nature and properties of the inducible sodium-plus-potassium ion-dependent adenosine triphosphatase in the gills of eels (Anguilla anguilla) adapted to fresh water and sea water.

Sargent J R, Thompson A J

Biochem J. 1974 Oct;144(1):69-75. doi: 10.1042/bj1440069.

Abstract

Gill tissue from eels adapted to fresh water or to sea water was disrupted in 0.32m-sucrose containing 0.1% (w/v) sodium deoxycholate and the subcellular distribution of (Na(+)+K(+))-dependent adenosine triphosphatase was determined. 2. About 70% of the recovered enzyme was in a fraction sedimenting between 225000g(av.)-min and 6000000g(av.)-min; the specific activities of enzymes from tissues of freshwater and seawater eels were 16 and 51 mumol of phosphate/h per mg of protein respectively. 3. The enzymes from gills of freshwater and seawater eels were indistinguishable on the basis of a number of parameters. These included phosphorylation by [gamma-(32)P]ATP, the binding of [(3)H]ouabain, the extent to which bound [(3)H]ouabain was displaced by increasing concentrations of KCl and pH optima. 4. Electrophoresis on polyacrylamide gels in sodium dodecyl sulphate showed that enzyme preparations from both sources had an identical number of protein components. 5. The higher specific activity of (Na(+)+K(+))-dependent adenosine triphosphatase from tissue of seawater eels was accompanied by increased amounts of two protein components. One of these proteins retained (32)P after treatment of the enzyme with [gamma-(32)P]ATP and had mol.wt. 97000; the other component was a glycoprotein with mol.wt. approx. 46000. 6. The results are discussed in terms of the nature of the transepithelial NaCl pumps in the gills of freshwater and seawater fish.

摘要

将适应淡水或海水的鳗鱼鳃组织在含有0.1%（w/v）脱氧胆酸钠的0.32m蔗糖溶液中匀浆，然后测定（Na⁺+K⁺）依赖性三磷酸腺苷酶的亚细胞分布。2. 回收的酶约70%存在于225000g（平均）-分钟至6000000g（平均）-分钟之间沉降的组分中；淡水鳗鱼和海水鳗鱼组织中酶的比活性分别为每毫克蛋白质16和51微摩尔磷酸盐/小时。3. 根据一些参数，淡水鳗鱼和海水鳗鱼鳃中的酶无法区分。这些参数包括[γ-（³²）P]ATP的磷酸化、[（³）H]哇巴因的结合、随着KCl浓度增加结合的[（³）H]哇巴因被置换的程度以及最适pH值。4. 在十二烷基硫酸钠聚丙烯酰胺凝胶上进行电泳表明，两种来源的酶制剂具有相同数量的蛋白质组分。5. 海水鳗鱼组织中（Na⁺+K⁺）依赖性三磷酸腺苷酶较高的比活性伴随着两种蛋白质组分含量的增加。其中一种蛋白质在用[γ-（³²）P]ATP处理酶后保留了（³²）P，分子量为97000；另一种组分是分子量约为46000的糖蛋白。6. 根据淡水鱼和海水鱼鳃中跨上皮NaCl泵的性质对结果进行了讨论。