An improved method for isolation of basolateral membranes from rat kidney.

An improved method for isolation of basolateral membranes from rat kidney.
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一种从大鼠肾脏中分离基底外侧膜的改进方法。

DOI:
10.1016/0005-2736(80)90516-7
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发表时间:
1980
期刊:
Biochimica et biophysica acta
影响因子:
--
通讯作者:
Segal,S
Segal,S
中科院分区:
--
文献类型:
--
作者:
Reynolds,RA;Wald,H;McNamara,PD;Segal,S

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我们已经描述了一种方法,采用差速离心和自由流电泳的分离基底外侧膜从大鼠肾皮质组织。膜制备的纯度进行了评估,通过测定酶的成分,并证实了d-和l-葡萄糖,l-脯氨酸和l-谷氨酰胺的运输特性的观察。(Na ++ K+)-ATP酶的比活性是皮层匀浆的15.4倍,相对纯化度是碱性磷酸酶的25倍。l-葡萄糖显示无Na+-梯度依赖性,并用作扩散的量度。摄取D-葡萄糖以及L-脯氨酸遵循相同的时间过程中存在或不存在的Na+梯度。只有与L-谷氨酰胺的摄取刺激的存在下的Na+梯度。我们的数据表明,我们成功地分离了基底外侧膜,它没有刷状缘污染,产率上级,比以前报道的更容易。
We have described a method using differential centrifugation and free-flow electrophoresis for the isolation of basolateral membranes from rat renal cortical tissue. Purity of the membrane preparation was assessed by determination of enzymic constituents and substantiated by observation of transport characteristics for d-and l-glucose, l-proline and l-glutamine. Specific activity of (Na++ K+)-ATPase in the pooled electrophoretic fractions was 15.4 times that in the cortical homogenate, and the relative purification of (Na++ K+)-ATPase compared to alkaline phosphatase was 25-fold. l-Glucose showed no Na+-gradient dependence and was used as a measure of diffusion. Uptake of d-glucose as well as l-proline followed the same time course in the presence or absence of an Na+ gradient. Only with l-glutamine was uptake stimulated by the presence of an Na+ gradient. Our data indicate we have sucessfully isolated basolateral membranes which are free of brush border contamination in superior yield and with greater facility than previously reported.
L-脯氨酸和 D-葡萄糖在肾皮质腔内(刷状缘)和腔内(基底外侧)膜囊泡中的转运。
DOI: --
发表时间: 1977
期刊: Biochemical and Biophysical Research Communications - BBRC
影响因子: --
作者:
E. Slack;T. Liang;B. Sacktor
通讯作者: B. Sacktor
钠加钾在肾脏中转运三磷酸腺苷酶活性。
DOI: 10.1002/jcp.1030620110
发表时间: 1963
影响因子: --
作者:
C. R. Kinsolving;R. Post;D. Beaver
通讯作者: D. Beaver
DOI: --
发表时间: 1969
影响因子: 4.8
作者:
D. Fitzpatrick;G. Davenport;L. Forte;E. Landon
通讯作者: E. Landon
DOI: 10.1016/0005-2744(68)90176-9
发表时间: 1968-01-01
期刊: BIOCHIMICA ET BIOPHYSICA ACTA
影响因子: --
作者:
JORGENSEN, PL
通讯作者: JORGENSEN, PL
DOI: 10.1083/jcb.54.2.232
发表时间: 1972-08
期刊: The Journal of cell biology
影响因子: --
作者:
Heidrich HG;Kinne R;Kinne-Saffran E;Hannig K
通讯作者: Hannig K