Transfer of glucagon receptor from liver membranes to a foreign adenylate cyclase by a membrane fusion procedure.
Transfer of glucagon receptor from liver membranes to a foreign adenylate cyclase by a membrane fusion procedure.
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通过膜融合程序将胰高血糖素受体从肝膜转移至外源腺苷酸环化酶。
DOI：
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发表时间：
1979
影响因子：
11.1
通讯作者：
M. Schramm
中科院分区：
文献类型：
--
作者：
M. Schramm
以前的工作已经证明，当红细胞上的β-肾上腺素能受体与组织培养细胞的腺苷酸环化酶[ATP焦磷酸裂解酶（环化），EC 4.6.1.1]通过仙台病毒融合时，这两种细胞发生偶联。这一发现对一般动物组织、膜制剂和肽类激素受体的有效性迄今尚未评估。可用的融合程序仅对来自组织培养的某些完整细胞和红细胞有效。在目前的工作中，膜融合的方法，导致胰高血糖素受体从纯化的大鼠肝膜转移到朋友红白血病细胞，甚至直接转移到膜部分制备的朋友细胞成为可行的。因此，可以得出结论，正常组织膜中的肽激素受体具有与红细胞中β-肾上腺素能受体相似的性质：它作为可分离的独立单位存在于膜中，可以容易地与外源细胞的腺苷酸环化酶偶联。膜融合过程的效率是由于聚乙二醇、磷脂、硬脂胺和ATP在盐介质中的联合作用。该方法有望适用于各种细胞和组织的膜，它可能被用来分析激素受体和腺苷酸环化酶系统的故障状态，通过转移到它们各自的对应物在正常细胞膜。膜成分的生物化学杂交研究不限于腺苷酸环化酶的激素激活。借助于膜融合方法，该方法可应用于生物膜中任何可解离的多组分体系。
Previous work had demonstrated the coupling of a beta-adrenergic receptor on an erythrocyte with the adenylate cyclase [ATP pyrophosphate-lyase (cyclizing), EC 4.6.1.1] of a tissue culture cell when the two cells were fused by Sendai virus. The validity of this finding for animal tissues in general, for membrane preparations, and for peptide hormone receptors could hitherto not be assessed. Available fusion procedures worked efficiently only with certain intact cells from tissue culture and with erythrocytes. In the present work a membrane fusion method was developed that causes the transfer of the glucagon receptor from purified rat liver membranes to Friend erythroleukemia cells; even direct transfer to a membrane fraction prepared from Friend cells became feasible. It can therefore be concluded that a peptide hormone receptor in a normal tissue membrane has properties similar to those demonstrated for a beta-adrenergic receptor in an erythrocyte: it exists in the membrane as a dissociable independent unit that can readily couple with the adenylate cyclase of a foreign cell. The efficiency of the membrane fusion procedure is due to the combined action of polyethylene glycol, phospholipids, stearylamine, and ATP in a salt medium. The method promises to be applicable to membranes of various cells and tissues, and it can probably be used to analyze hormone receptors and adenylate cyclase systems in states of malfunction by transfer to their respective counterpart in a normal cell membrane. Studies in biochemical hybridization of membrane components need not be limited to hormone activation of adenylate cyclase. With the aid of the membrane fusion method, this approach could be applied to any dissociable multicomponent system in biological membranes.