G蛋白偶联受体的异源表达：从大规模生产和纯化角度比较表达系统
Heterologous expression of G-protein-coupled receptors: comparison of expression systems from the standpoint of large-scale production and purification.
作者信息
Sarramegna V, Talmont F, Demange P, Milon A
机构信息
Institut de Pharmacologie et de Biologie Structurale, Toulouse, Cedex 4, France.
出版信息
Cell Mol Life Sci. 2003 Aug;60(8):1529-46. doi: 10.1007/s00018-003-3168-7.
Abstract
G-protein-coupled receptors (GPCRs) are of prime importance for cell signal transduction mechanisms and are the target of many current and potential drugs. However, structural data on these membrane proteins is still scarce because of their low natural abundance and the low efficiency of most of the expression systems currently available. This review presents the most important expression systems currently employed for heterologous expression of GPCRs; Escherichia coli, yeast, insect cells and mammalian cells. After briefly recalling the specificity, advantages and limitations of each system, particular emphasis is put on the quantitative comparison of these expression systems in terms of overall expression yield, and on the influence of various factors (primary sequence, origin, cell type, N- and C-terminal tags) on the results.
摘要
G蛋白偶联受体（GPCRs）对于细胞信号转导机制至关重要，并且是许多现有及潜在药物的作用靶点。然而，由于这些膜蛋白天然丰度低以及目前大多数可用表达系统效率低下，关于它们的结构数据仍然稀缺。本综述介绍了目前用于GPCRs异源表达的最重要的表达系统；大肠杆菌、酵母、昆虫细胞和哺乳动物细胞。在简要回顾每个系统的特异性、优点和局限性之后，特别强调了这些表达系统在整体表达产量方面的定量比较，以及各种因素（一级序列、来源、细胞类型、N端和C端标签）对结果的影响。
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本文引用的文献
1
Guidelines for cardiac pacing and cardiac resynchronization therapy: The Task Force for Cardiac Pacing and Cardiac Resynchronization Therapy of the European Society of Cardiology. Developed in collaboration with the European Heart Rhythm Association.心脏起搏与心脏再同步治疗指南：欧洲心脏病学会心脏起搏与心脏再同步治疗工作组。与欧洲心律协会合作制定。
Eur Heart J. 2007 Sep;28(18):2256-95. doi: 10.1093/eurheartj/ehm305. Epub 2007 Aug 28.
2
High prevalence of sleep apnea syndrome in patients with long-term pacing: the European Multicenter Polysomnographic Study.长期起搏患者睡眠呼吸暂停综合征的高患病率：欧洲多中心多导睡眠图研究
Circulation. 2007 Apr 3;115(13):1703-9. doi: 10.1161/CIRCULATIONAHA.106.659706. Epub 2007 Mar 12.
3
Association of nocturnal arrhythmias with sleep-disordered breathing: The Sleep Heart Health Study.夜间心律失常与睡眠呼吸障碍的关联：睡眠心脏健康研究
Am J Respir Crit Care Med. 2006 Apr 15;173(8):910-6. doi: 10.1164/rccm.200509-1442OC. Epub 2006 Jan 19.
4
Atrial overdrive pacing for the obstructive sleep apnea-hypopnea syndrome.用于阻塞性睡眠呼吸暂停低通气综合征的心房超速起搏
N Engl J Med. 2005 Dec 15;353(24):2568-77. doi: 10.1056/NEJMoa050610.
5
[Guidelines for heart pacemaker therapy].[心脏起搏器治疗指南]
Z Kardiol. 2005 Oct;94(10):704-20. doi: 10.1007/s00392-005-0269-3.
6
Severe bradyarrhythmias in patients with sleep apnoea: the effect of continuous positive airway pressure treatment: a long-term evaluation using an insertable loop recorder.睡眠呼吸暂停患者的严重缓慢性心律失常：持续气道正压通气治疗的效果：使用植入式环形记录仪的长期评估
Eur Heart J. 2004 Jun;25(12):1070-6. doi: 10.1016/j.ehj.2004.04.017.
7
Images in cardiovascular medicine. Malignant vasovagal syncope.心血管医学影像。恶性血管迷走性晕厥。
Circulation. 2003 Jun 17;107(23):2987-8. doi: 10.1161/01.CIR.0000068033.19278.F1.
8
Relationship among the severity of sleep apnea syndrome, cardiac arrhythmias, and autonomic imbalance.睡眠呼吸暂停综合征的严重程度、心律失常和自主神经失衡之间的关系。
Pacing Clin Electrophysiol. 2003 Mar;26(3):669-77. doi: 10.1046/j.1460-9592.2003.00116.x.
9
Cardiac rhythm disturbances in the obstructive sleep apnea syndrome: effects of nasal continuous positive airway pressure therapy.阻塞性睡眠呼吸暂停综合征中的心律失常：经鼻持续气道正压通气治疗的效果
Chest. 2000 Sep;118(3):591-5. doi: 10.1378/chest.118.3.591.
10
Outcome of patients with sleep apnea-associated severe bradyarrhythmias after continuous positive airway pressure therapy.持续气道正压通气治疗后睡眠呼吸暂停相关严重缓慢性心律失常患者的结局
Am J Cardiol. 2000 Sep 15;86(6):688-92, A9. doi: 10.1016/s0002-9149(00)01055-9.
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