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通过膜融合程序将胰高血糖素受体从肝细胞膜转移至外源腺苷酸环化酶。

Transfer of glucagon receptor from liver membranes to a foreign adenylate cyclase by a membrane fusion procedure.

作者信息

Schramm M

出版信息

Proc Natl Acad Sci U S A. 1979 Mar;76(3):1174-8. doi: 10.1073/pnas.76.3.1174.

DOI:10.1073/pnas.76.3.1174
PMID:220608
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC383212/
Abstract

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.

摘要

先前的研究表明，当用仙台病毒使红细胞上的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]融合时，二者能够偶联。此前，这一发现对于一般动物组织、膜制剂以及肽激素受体的有效性尚无法评估。现有的融合方法仅对某些来自组织培养的完整细胞和红细胞有效。在本研究中，开发了一种膜融合方法，可使纯化的大鼠肝细胞膜上的胰高血糖素受体转移至弗氏红白血病细胞；甚至直接转移至由弗氏细胞制备的膜组分也变得可行。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-肾上腺素能受体类似的特性：它以可解离的独立单位存在于膜中，能够轻易地与异源细胞的腺苷酸环化酶偶联。膜融合程序的效率归因于聚乙二醇、磷脂、硬脂胺和ATP在盐介质中的联合作用。该方法有望应用于各种细胞和组织的膜，并且可能用于通过转移至正常细胞膜中的相应对应物来分析功能异常状态下的激素受体和腺苷酸环化酶系统。膜组分的生化杂交研究不必局限于腺苷酸环化酶的激素激活。借助膜融合方法，这种方法可应用于生物膜中任何可解离的多组分系统。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/ea1f/383212/e5330a05b688/pnas00003-0173-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/ea1f/383212/e5330a05b688/pnas00003-0173-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/ea1f/383212/e5330a05b688/pnas00003-0173-a.jpg

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