Chromatographic resolution of insulin receptor from insulin-sensitive D-glucose transporter of adipocyte plasma membranes.

Chromatographic resolution of insulin receptor from insulin-sensitive D-glucose transporter of adipocyte plasma membranes.
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脂肪细胞质膜胰岛素敏感 D-葡萄糖转运蛋白的胰岛素受体的色谱分离。

DOI:
10.1021/bi00504a036
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发表时间:
1981
期刊:
影响因子:
2.9
通讯作者:
Czech,MP
Czech,MP
中科院分区:
生物学3区
文献类型:
--
作者:
Carter-Su,C;Pilch,PF;Czech,MP

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Christin Carter-Su,* Paul F. Pilch和Michael P. Czech摘要:为了检验转运蛋白可能由受体多肽组成的假设,对亲和标记的脂肪细胞胰岛素受体和胰岛素敏感的己糖转运蛋白的色谱特性进行了评价。通过使用交联剂辛二酸二琥珀酰亚胺酯[Pilch,P.F.,& Czech,M. P.(1979)J.Biol.Chem.254,3375-3381],用胆酸钠溶解膜,并在羟基磷灰石色谱法后分解成三个蛋白质峰。当重组到磷脂囊泡中时,第一个或空隙体积峰含有10-20%的蛋白质,没有受体结合的[125 I]胰岛素,也没有细胞松弛素B敏感的D-葡萄糖转运活性。第二个主峰含有约一半的蛋白质和多达87%的亲和标记受体,但没有己糖转运活性。第三个峰含有30-40%的蛋白质,13-20%的-A。尽管在了解激素如何与受体蛋白相互作用以及激素-受体相互作用导致细胞功能变化方面取得了重大进展,但对膜结合激素受体及其效应系统的空间关系知之甚少。可获得此类信息的两种激素受体效应系统是乙酰胆碱和β-肾上腺素能受体效应系统。Huganir等人(1979)最近报道乙酰胆碱依赖性Na+通道似乎是与乙酰胆碱受体相同的蛋白质复合物的一部分。的
Christin Carter-Su,* Paul F. Pilch, and Michael P. Czech abstract: The chromatographic properties of the affinity-labeled adipocyte insulin receptor and the insulin-sensitive hexose transporter were evaluated in order to test the hypothesis that the transportermight be composed of receptor polypeptides.[I25I] Insulin was linked specifically and cova-lently to the high-affinity insulinreceptor in dimethylmaleic anhydride extracted rat adipocyte plasma membranes by using the cross-linking reagent disuccinimidyl suberate [Pilch, P. F., & Czech, M. P.(1979) J. Biol. Chem. 254, 3375-3381], The membranes were solubilized with sodium cholate and resolved into three peaks of protein following hydroxylapatite chro-matography. The first or void volume peak contained 10-20% of the protein, no receptor-bound [125I] insulin, and no cytochalasin B sensitive D-glucose transport activity when recon-stituted into phospholipid vesicles. The second major peak contained about half of the protein and as much as 87% of the affinity-labeled receptor but no hexose transport activity. The third peak contained 30-40% of the protein, 13-20% of-A-lthough significant advances have been made in understanding how hormones interact with receptor proteins and what changes in cell function are brought about as a result of hormone-receptorinteractions, little is known about the spatial relationship of membrane-bound hormone receptors and their effector systems. Two of the hormone receptoreffector systems for which such information is available are the acetylcholine and/3-adrenergic receptor-effector systems. Huganir et al.(1979) have recently reported that the ace-tylcholine-dependent Na+ channel appears to be part of the same protein complex as the acetylcholine receptor. The
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