Application of immunochemical methods to the identification and characterization of rat kidney inactive renin.
Application of immunochemical methods to the identification and characterization of rat kidney inactive renin.
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免疫化学方法在大鼠肾脏失活肾素鉴定和表征中的应用。
DOI：
10.1161/01.hyp.7.2.236
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发表时间：
1985
期刊：
影响因子：
--
通讯作者：
Inagami,T
中科院分区：
文献类型：
--
作者：
Takii,Y;Figueiredo,AF;Inagami,T
由于组织中或在均质化和纯化过程中无活性的酶原会迅速蛋白水解转化为其活性形式，因此很难识别肾脏中的无活性原。免疫化学方法，蛋白质印迹，直接放射免疫分析，免疫亲和层析分离和鉴定大鼠肾肾素和原肾素，并确定其分子量没有完全纯化。在家兔中制备了抗纯大鼠肾素的抗血清。免疫双扩散、酶活性抑制和竞争性放射免疫分析表明，抗血清与大鼠肾素发生特异性反应。抗大鼠肾素IgG与纯化的人肾素或大鼠脾或肾组织蛋白酶D无交叉反应。IgG显示出对失活的肾素以及活性酶的结合亲和力。由胃蛋白酶抑制剂-琼脂糖凝胶，IgG-琼脂糖凝胶和Affi-Gel Blue组成的亲和层析的组合允许从大鼠肾提取物中的活性肾素快速和完全分离非活性肾素。无论是非活性还是活性的肾素制剂都没有表现出对血红蛋白作为底物的乙酰化蛋白酶活性。通过凝胶过滤法估计无活性的肾素的表观分子量为50，000。在十二烷基硫酸钠（SDS）聚丙烯酰胺凝胶中电泳部分纯化的无活性肾素，然后将蛋白质转移到硝酸纤维素片上，并用抗肾素IgG进行免疫化学染色，结果显示分子量为48，000的单一蛋白质条带。胰蛋白酶对失活的肾素的激活伴随着48，000-道尔顿的天然蛋白质还原成39，000-道尔顿的蛋白质，如SDS聚丙烯酰胺凝胶电泳和transblotting所测定的。（250字处删节）
Identification of inactive prorenin in the kidney has been difficult due to rapid proteolytic conversion of the inactive zymogen to its active form in the tissue or during homogenization and purification. Immunochemical methods, Western blotting, direct radioimmunoassay, and immunoaffinity chromatography were used to isolate and identify rat kidney renin and prorenin and to determine their molecular weights without complete purification. Antisera to pure rat renin were raised in rabbits. A specific reaction between the antisera and rat renin was demonstrated by double immunodiffusion, inhibition of enzyme activity, and competitive radioimmunoassay. The anti-rat renin IgG did not cross-react with purified human renin or rat spleen or kidney cathepsin D. The IgG showed binding affinity to both inactive renin as well as active enzyme. A combination of affinity chromatographies consisting of pepstatin-Sepharose, IgG-Sepharose, and Affi-Gel Blue permitted rapid and complete separation of inactive renin from active renin in rat kidney extract. Neither inactive nor active renin preparations exhibited aspartyl protease activity on hemoglobin used as substrate. The apparent molecular weight of inactive renin was estimated as 50,000 by gel filtration. Electrophoresis of partially purified inactive renin in sodium dodecyl sulfate (SDS) polyacrylamide gel followed by transblotting of proteins to a nitrocellulose sheet and immunochemical staining with anti-renin IgG showed a single protein band with a molecular weight of 48,000. Activation of inactive renin by trypsin was accompanied by the reduction of the 48,000-dalton native protein to a 39,000-dalton protein as determined by the SDS polyacrylamide gel electrophoresis and the transblotting.(ABSTRACT TRUNCATED AT 250 WORDS)
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影响因子：
7.8
作者：
H. Reggio;G. Palade
通讯作者：
G. Palade
雪貂气管腺35SO4-大分子分泌的神经调节。
DOI：
10.1152/jappl.1984.57.2.457
发表时间：
1984
期刊：
Journal of applied physiology: respiratory, environmental and exercise physiology
影响因子：
--
作者：
Borson,DB;Charlin,M;Gold,BD;Nadel,JA
通讯作者：
Nadel,JA
雪貂气管粘膜下腺中的肾上腺素能和胆碱能受体：放射自显影定位。
DOI：
--
发表时间：
1983
期刊：
European journal of respiratory diseases. Supplement
影响因子：
--
作者：
C. Basbaum;P. Barnes;M. Grillo;J. Widdicombe;J. Nadel
通讯作者：
J. Nadel
人类气管支气管腺粘液细胞产生的酸性糖蛋白的组织化学类型
DOI：
10.1002/path.1710980402
发表时间：
1969
期刊：
The Journal of Pathology
影响因子：
--
作者：
D. Lamb;Lynne M. Reid
通讯作者：
Lynne M. Reid
拟交感神经药物在体外刺激人支气管分泌糖蛋白的输出。
影响因子：
6
作者：
R. Phipps;Isobel P. Williams;P. Richardson;J. Pell;R. Pack;N. Wright
通讯作者：
N. Wright