Structure and function in rhodopsin: High-level expression of rhodopsin with restricted and homogeneous N-glycosylation by a tetracycline-inducible N-acetylglucosaminyltransferase I-negative HEK293S…

By P. Reeves, N. Callewaert, R. Contreras, H. Khorana. 2002

623 citations recorded by the source corpus.

Subfields: Cellular and Molecular Neuroscience, Molecular Biology, Surgery

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In Cellular and Molecular Neuroscience

1. D. Virshup

In Molecular Biology

1. P. Talbot
2. A. Wong
3. M. Desforges
4. Jonathan Kim
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6. Dongxia Zhou
7. John L. Rubinstein
8. G. Parisi

Citations in the corpus

• Structural basis of WLS/Evi-mediated Wnt transport and secretion (2020, methodology): “WNT8A and WLS were co-expressed in HEK293S GnTI− cells (Reeves et al., 2002) by co-transduction of two separate baculoviruses prepared with the BacMam system (Goehring et al.”
• The human coronavirus HCoV-229E S-protein structure and receptor binding (2019, methodology): “Stable cell lines were generated by co-transfecting HEK293S-GnT1(-/-) cells (ATCC Cat# CRL-3022, RRID: CVCL_A785) (Reeves et al., 2002) with each of the RBD plasmids and both a piggyBac transposase helper plasmid and an rtTA helper plasmid.”
• Structural activation of pro-inflammatory human cytokine IL-23 by cognate IL-23 receptor enables recruitment of the shared receptor IL-12Rβ1 (2017, background): “…IL-23, and the ectodomains of IL-23R (residues 1–317) and IL-12R β 1 (residues 1– 540) (Figure 1A), via inducible stable expression and secretion in HEK293S MGAT1 −/− cells, which limit N-linked glycosylation of proteins to Man 5 GlcNAc 2 glycan trees (Reeves et al., 2002; Verstraete et al., 2011).”
• Structure of HIV-1 gp120 V1/V2 domain with broadly neutralizing antibody PG9 (2011, neutral): “Scaffold-V1/V2 heterogeneity was apparent after expression in GnTI −/− cell”
• Structures of Aplysia AChBP complexes with nicotinic agonists and antagonists reveal distinctive binding interfaces and conformations (2005, background): “However, expression from a glycosylation-deficient cell line (Reeves et al, 2002) resulted in a homogenous protein with a shorter glycan chain as assessed by electrophoresis (Figure 1A) and mass spectrometry analyses, yielding crystals suitable for structural studies.”
• BacMam system for high-level expression of recombinant soluble and membrane glycoproteins for structural studies. (2008, methodology): “The 293 GnTI− cell line developed by Khorana and collegues is a HEK293S derived cell line that lacks the capability to process N-linked glycans on glycoproteins from the high mannose to the complex mature form due to inactivation of both alleles of the GnTI gene [7].”