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从哺乳动物大脑皮层纯化的γ-氨基丁酸A受体的分子大小。

Molecular size of the gamma-aminobutyric acidA receptor purified from mammalian cerebral cortex.

作者信息

Mamalaki C, Barnard E A, Stephenson F A

机构信息

MRC Molecular Neurobiology Unit, Medical Research Council Centre, Cambridge, England.

出版信息

J Neurochem. 1989 Jan;52(1):124-34. doi: 10.1111/j.1471-4159.1989.tb10906.x.

Abstract

The hydrodynamic behaviour of both the soluble and purified gamma-aminobutyric acidA (GABAA) receptor of bovine or rat cerebral cortex has been investigated in solution in Triton X-100 or in 3-[(3-cholamidopropyl)-dimethylammonio]-1-propanesulphonate (CHAPS). In all the hydrodynamic separations made, it was found that the binding activities for GABA, benzodiazepine, and (where detectable) t-butylbicyclophosphorothionate comigrated. Conditions were established for gel exclusion chromatography and for sucrose density gradient velocity sedimentation that maintain the GABAA receptor in a nonaggregated form. Using these conditions, the molecular weight of the bovine GABAA receptor in the above-mentioned detergents was calculated using the H2O/2H2O method. A value of Mr 230,000-240,000 was calculated for the bovine pure GABAA receptor purified in sodium deoxycholate/Triton X-100 media. A value of Mr 284,000-290,000 was calculated for the nonaggregated bovine or rat cortex receptor in CHAPS, but the Stokes radius is smaller in the latter than in the former medium and the detergent binding in CHAPS is underestimated. Thus the deduced Mr, 240,000, is the best estimate by this method.

摘要

已在Triton X-100溶液或3-[(3-胆酰胺丙基)-二甲基铵]-1-丙烷磺酸盐(CHAPS)中,对牛或大鼠大脑皮层可溶性及纯化的γ-氨基丁酸A(GABAA)受体的流体动力学行为进行了研究。在所有进行的流体动力学分离中,发现GABA、苯二氮卓类药物以及(若可检测到)硫代丁基双环磷酰硫酯的结合活性共同迁移。建立了凝胶排阻色谱法和蔗糖密度梯度速度沉降法的条件,以保持GABAA受体呈非聚集形式。使用这些条件,采用H2O/2H2O方法计算了上述去污剂中牛GABAA受体的分子量。在脱氧胆酸钠/Triton X-100介质中纯化的牛纯GABAA受体,计算得出的分子量为230,000 - 240,000。在CHAPS中,非聚集的牛或大鼠皮层受体计算得出的分子量为284,000 - 290,000,但后者的斯托克斯半径比前者介质中的小,且CHAPS中的去污剂结合被低估。因此,通过该方法推断出的240,000是最佳估计值。

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