Isolation and characterization of plasma membranes from bovine carotid arteries.
Isolation and characterization of plasma membranes from bovine carotid arteries.
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牛颈动脉质膜的分离和表征。
DOI：
10.1152/ajpcell.1986.250.1.c65
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发表时间：
1986
期刊：
影响因子：
--
通讯作者：
Bhalla,RC
中科院分区：
文献类型：
--
作者：
Sharma,RV;Bhalla,RC
A plasma membrane fraction from bovine carotid arteries has been isolated by extraction of a crude microsomal fraction with a low-ionic-strength buffer containing ATP and Ca2+. This step was followed by sucrose-density-gradient centrifugation in the presence of 0.6 M KCl. The plasma membrane vesicles were enriched 60- to 80-fold in Na+-K+-adenosinetriphosphatase, 5'-nucleotidase, and phosphodiesterase I activities. The final yields of these marker enzymes were 12-18% of the total activities in the postnuclear supernatant, and the protein yield was 100-120 micrograms/g wet wt of carotid arteries. Contamination of the plasma membrane fraction by mitochondria and sarcoplasmic reticulum was low as judged by low activities of succinate--cytochrome-c reductase and NADPH--cytochrome-c reductase, respectively. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis and immunoprecipitation with smooth muscle-specific actin antibodies showed that the plasma membrane fraction was substantially free from myosin and actin contamination. The plasma membrane vesicles accumulated Ca2+ in the presence of ATP, and the accumulation was increased by calmodulin. Ca2+ accumulated in the presence or absence of calmodulin could be released almost completely from the vesicles by the addition of the Ca2+ ionophore A23187 but not by ethyleneglycol-bis(beta-aminoethylether)-N,N'-tetraacetic acid, indicating that Ca2+ uptake in the presence of ATP is intravesicular. The effects of phosphate and oxalate on Ca2+ uptake in the plasma membranes were different from one another. Phosphate increased Ca2+ uptake in a concentration- and time-dependent manner, and the increase in Ca2+ uptake could be observed as early as 1 min. On the other hand, oxalate at concentrations up to 5 mM did not increase Ca2+ uptake significantly during the 30-min incubation. These plasma membranes can prove useful for the study of ion transport across plasma membranes, hormone binding, characterization of calcium channels, and preparation of antibodies against plasma membrane proteins.
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长春新碱与小牛脑微管蛋白的相互作用。
DOI：
--
发表时间：
1983
期刊：
The Journal of biological chemistry
影响因子：
--
作者：
Prakash,V;Timasheff,SN
通讯作者：
Timasheff,SN
微管蛋白在中性 pH 值下的老化。
影响因子：
5.6
作者：
Prakash,V;Timasheff,SN
通讯作者：
Timasheff,SN
影响因子：
2.9
作者：
R. P. Frigon;S. N. Timasheff
通讯作者：
S. N. Timasheff
在存在和不存在配体的情况下，分析超速离心机中双峰的分辨率。
影响因子：
2.9
作者：
Prakash,V;Timasheff,SN
通讯作者：
Timasheff,SN
影响因子：
2.9
作者：
R. P. Frigon;S. N. Timasheff
通讯作者：
S. N. Timasheff