Insulin receptors from guinea pig liver and brain: structural and functional studies.
Insulin receptors from guinea pig liver and brain: structural and functional studies.
复制标题
豚鼠肝脏和大脑的胰岛素受体：结构和功能研究。
作者：
W. Lowe;D. Leroith
我们研究了豚鼠肝脏和大脑中胰岛素受体的结构和功能特征。与肝和脑粗膜制剂的结合是时间、温度和pH依赖性的。对肝粗膜制剂的最大特异性结合为16.4 +/- 0.5%，对脑粗膜制剂的最大特异性结合为10.4 +/- 1.8%。特异性研究表明，在肝脏和大脑中，胰岛素受体与鸡胰岛素的典型亲和力高于猪胰岛素，高于人胰岛素原，高于去肽胰岛素。抗胰岛素受体抗血清抑制[125I]胰岛素与肝和脑粗膜制剂的结合。肝脏和脑胰岛素受体与[125I]胰岛素亲和交联后，在还原条件下进行电泳，发现标记蛋白（α亚基）的表观摩尔重量在肝脏为136,000，在脑为121,000。用内糖苷酶H和内糖苷酶F处理肝脏和脑受体可增加α -亚基的电泳迁移率。[125I]通过结合和十二烷基硫酸钠-聚丙烯酰胺凝胶电泳证实，肝脏和脑的胰岛素交联受体以类似的方式吸附在小麦胚芽凝集素柱上并从其洗脱。在肝脏和脑的可溶性凝集素纯化受体制剂中，胰岛素刺激β亚基和外源性底物的磷酸化。当通过外源性底物磷酸化测量脑和肝制剂的δ激酶活性时，将其归一化为制剂的最大结合与游离比，脑中的δ激酶活性比肝中的δ激酶活性高约4倍。这些研究表明，先前在大鼠中证明的脑和肝胰岛素受体α亚基之间的差异也存在于豚鼠中。
We studied the structural and functional characteristics of insulin receptors from guinea pig liver and brain. Binding to crude membrane preparations of liver and brain was time, temperature, and pH dependent. Maximal specific binding to liver crude membrane preparations was 16.4 +/- 0.5%, and that to brain crude membrane preparations was 10.4 +/- 1.8%. Specificity studies demonstrated typical affinities for insulin receptors with chicken insulin greater than porcine insulin greater than human proinsulin greater than desoctapeptide insulin in both liver and brain. Antiinsulin receptor antiserum inhibited binding of [125I]insulin to both liver and brain crude membrane preparations. Electrophoresis performed under reducing conditions after affinity cross-linking of liver and brain insulin receptors with [125I]insulin revealed labeled proteins (alpha-subunit) with apparent mol wt of 136,000 in liver and 121,000 in brain. Treatment of liver and brain receptors with both endoglycosidase H and endoglycosidase F increased the electrophoretic mobility of the alpha-subunit. [125I]Insulin cross-linked receptors from both liver and brain adsorbed to and eluted from wheat germ agglutinin columns in a similar manner, as demonstrated by binding and sodium dodecyl sulfate-polyacrylamide gel electrophoresis. In solubilized lectin-purified receptor preparations from liver and brain, insulin stimulated the phosphorylation of the beta-subunit and exogenous substrates. When the delta-kinase activity, as measured by exogenous substrate phosphorylation, of the brain and liver preparations was normalized to the maximal bound to free ratio of the preparations, the delta-kinase activity was about 4-fold greater in brain than in liver. These studies suggest that the differences between brain and liver insulin receptor alpha-subunits previously demonstrated in rats are also present in guinea pigs.
登录
查看更多内容
DOI：
10.1016/s0021-9258(18)32085-4
发表时间：
1983-07
期刊：
The Journal of biological chemistry
影响因子：
--
作者：
K. Heidenreich;N. Zahniser;P. Berhanu;D. Brandenburg;J. Olefsky
通讯作者：
K. Heidenreich;N. Zahniser;P. Berhanu;D. Brandenburg;J. Olefsky
DOI：
10.1152/ajpendo.1985.248.1.e115
发表时间：
1985
期刊：
The American journal of physiology
影响因子：
--
作者：
Haskell,JF;Meezan,E;Pillion,DJ
通讯作者：
Pillion,DJ
DOI：
10.1073/pnas.79.15.4540
发表时间：
1982-01-01
期刊：
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA-BIOLOGICAL SCIENCES
影响因子：
--
作者：
ELDER, JH;ALEXANDER, S
通讯作者：
ALEXANDER, S
人胎盘膜洗涤剂提取物中胰岛素刺激的胰岛素受体酪氨酸磷酸化。
DOI：
--
发表时间：
1982
期刊：
The Journal of biological chemistry
影响因子：
--
作者：
Avruch,J;Nemenoff,RA;Blackshear,PJ;Pierce,MW;Osathanondh,R
通讯作者：
Osathanondh,R
DOI：
10.1016/s0021-9258(18)34631-3
发表时间：
1982-05
期刊：
The Journal of biological chemistry
影响因子：
--
作者：
J. Massagué;M. Czech
通讯作者：
J. Massagué;M. Czech