PMID- 34085260
OWN - NLM
STAT- MEDLINE
DCOM- 20210708
LR  - 20210708
IS  - 1940-6029 (Electronic)
IS  - 1064-3745 (Linking)
VI  - 2268
DP  - 2021
TI  - Construction of Recombinant Cell Lines for GPCR Expression.
PG  - 43-60
LID - 10.1007/978-1-0716-1221-7_3 [doi]
AB  - Large-scale recombinant expression of G protein-coupled receptors (GPCRs) is 
      required for structure and function studies where there is a need for milligram 
      amounts of protein in pure form. Here we describe a procedure for the 
      construction of human embryonic kidney 293S (HEK293S) stable cell lines for 
      inducible expression of the gene encoding bovine rhodopsin. The HEK293S cell line 
      is particularly suitable for this application because of several favorable 
      properties as a recombinant host including: its ease of transfection, its 
      capacity for handling large amounts of protein cargo, and its ability to perform 
      the necessary co- and post-translational modifications required for correct 
      folding and processing of complex membrane proteins such as GPCRs. The procedures 
      described here will focus on the HEK293S GnTI(-) cell line, an HEK293S derivative 
      that is widely used for the production of glycoproteins modified homogeneously 
      with truncated N-glycans.
FAU - Reeves, Philip J
AU  - Reeves PJ
AD  - School of Life Sciences, University of Essex, Colchester, Essex, UK. 
      preeves@essex.ac.uk.
LA  - eng
PT  - Journal Article
PL  - United States
TA  - Methods Mol Biol
JT  - Methods in molecular biology (Clifton, N.J.)
JID - 9214969
RN  - 0 (Receptors, G-Protein-Coupled)
RN  - 0 (Recombinant Proteins)
RN  - 9009-81-8 (Rhodopsin)
SB  - IM
MH  - Animals
MH  - Cattle
MH  - Glycosylation
MH  - HEK293 Cells
MH  - Humans
MH  - *Protein Processing, Post-Translational
MH  - Receptors, G-Protein-Coupled/genetics/isolation & purification/*metabolism
MH  - Recombinant Proteins/chemistry/genetics/isolation & purification/*metabolism
MH  - Rhodopsin/genetics/isolation & purification/*metabolism
MH  - Transfection
OTO - NOTNLM
OT  - Calcium phosphate
OT  - Glycosylation
OT  - Sodium butyrate
OT  - Tetracycline
EDAT- 2021/06/05 06:00
MHDA- 2021/07/09 06:00
CRDT- 2021/06/04 07:19
PHST- 2021/06/04 07:19 [entrez]
PHST- 2021/06/05 06:00 [pubmed]
PHST- 2021/07/09 06:00 [medline]
AID - 10.1007/978-1-0716-1221-7_3 [doi]
PST - ppublish
SO  - Methods Mol Biol. 2021;2268:43-60. doi: 10.1007/978-1-0716-1221-7_3.