Loisel T P, Ansanay H, St-Onge S, Gay B, Boulanger P, Strosberg A D, Marullo S, Bouvier M
Département de Biochemie and Le Groupe de Recherche sur le Système Nerveux Autonome, Université de Montréal, Canada.
Nat Biotechnol. 1997 Nov;15(12):1300-4. doi: 10.1038/nbt1197-1300.
Expression in baculovirus-infected insect cells allows sufficient production of G-protein coupled receptor for structural studies. An important drawback of this expression system comes from the presence of unprocessed and biologically inactive receptors that have to be eliminated during receptor purification steps. We show that viral particles released from Sf9 cells infected with a recombinant baculovirus coding for the human beta 2-adrenergic receptor (beta 2AR) cDNA contain glycosylated and biologically active beta 2AR. In addition, post-translational modifications known to modulate receptor activity were found to occur in these particles.
在杆状病毒感染的昆虫细胞中表达能够产生足够量的G蛋白偶联受体用于结构研究。该表达系统的一个重要缺点是存在未加工且无生物活性的受体,这些受体在受体纯化步骤中必须被去除。我们发现,用编码人β2肾上腺素能受体(β2AR)cDNA的重组杆状病毒感染Sf9细胞后释放的病毒颗粒含有糖基化且具有生物活性的β2AR。此外,还发现已知可调节受体活性的翻译后修饰发生在这些颗粒中。