ArticlePloS one2013
Transient and stable expression of the neurotensin receptor NTS1: a comparison of the baculovirus-insect cell and the T-REx-293 expression systems.
Article in PloS one, 2013. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 4 papers.
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Who cites it
4 citing papers in PubMed, 14 citations in OpenAlex.
- ENPP1-Fc prevents mortality and vascular calcifications in rodent model of generalized arterial calcification of infancy.Nature communications · 2015Article
- MiRNA mimic screen for improved expression of functional neurotensin receptor from HEK293 cells.Biotechnology and bioengineering · 2015Article
- Engineering cells to improve protein expression.Current opinion in structural biology · 2014Review
- Better and faster: improvements and optimization for mammalian recombinant protein production.Current opinion in structural biology · 2014Review
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Authors and funding
5 authors at 4 institutions in 1 country.
Funding
Abstract
Nowadays, baculovirus-infected insect cells and tetracycline-inducible mammalian cell lines (T-REx-293) are intensively used for G protein-coupled receptor (GPCR) production for crystallography purposes. Here we constructed a suspension T-REx-293 cell line to stably express an engineered neurotensin receptor 1 (NTS1) mutant and we quantitatively compared this cell line with the transient baculovirus-insect cell system throughout a milligram-scale NTS1 expression and purification process. The two systems were comparable with respect to functional NTS1 expression levels and receptor binding affinity for the agonist [(3)H] neurotensin. However, NTS1 surface display on T-REx-293 cells determined by radio-ligand binding assays was 2.8 fold higher than that on insect cells. This work demonstrates two approaches for preparing milligram quantities of purified NTS1 suitable for structural studies and provides useful input to users in choosing and optimizing an appropriate expression host for other GPCRs.
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