scholarly journals Screening and large-scale expression of membrane proteins in mammalian cells for structural studies

2014 ◽  
Vol 9 (11) ◽  
pp. 2574-2585 ◽  
Author(s):  
April Goehring ◽  
Chia-Hsueh Lee ◽  
Kevin H Wang ◽  
Jennifer Carlisle Michel ◽  
Derek P Claxton ◽  
...  
Keyword(s):  
Membrane Proteins ◽  
Mammalian Cells ◽  
Large Scale ◽  
Structural Studies

scholarly journals Robotic large-scale application of wheat cell-free translation to structural studies including membrane proteins

2011 ◽  
Vol 28 (3) ◽  
pp. 239-249 ◽  
Author(s):  
Emily T. Beebe ◽  
Shin-ichi Makino ◽  
Akira Nozawa ◽  
Yuko Matsubara ◽  
Ronnie O. Frederick ◽  
...  
Keyword(s):  
Membrane Proteins ◽  
Large Scale ◽  
Structural Studies ◽  
Free Translation

scholarly journals Overexpression of membrane proteins in mammalian cells for structural studies

2012 ◽  
Vol 30 (1) ◽  
pp. 52-63 ◽  
Author(s):  
Juni Andréll ◽  
Christopher G. Tate
Keyword(s):  
Membrane Proteins ◽  
Mammalian Cells ◽  
Structural Studies

TMCC3 localizes at the three-way junctions for the proper tubular network of the endoplasmic reticulum

2019 ◽  
Vol 476 (21) ◽  
pp. 3241-3260
Author(s):  
Sindhu Wisesa ◽  
Yasunori Yamamoto ◽  
Toshiaki Sakisaka
Keyword(s):  
Endoplasmic Reticulum ◽  
Membrane Proteins ◽  
Membrane Protein ◽  
Mammalian Cells ◽  
Coiled Coil ◽  
Transmembrane Domains ◽  
Domain Family ◽  
Coiled Coil Domain ◽  
U2os Cells ◽  
Er Membrane

The tubular network of the endoplasmic reticulum (ER) is formed by connecting ER tubules through three-way junctions. Two classes of the conserved ER membrane proteins, atlastins and lunapark, have been shown to reside at the three-way junctions so far and be involved in the generation and stabilization of the three-way junctions. In this study, we report TMCC3 (transmembrane and coiled-coil domain family 3), a member of the TEX28 family, as another ER membrane protein that resides at the three-way junctions in mammalian cells. When the TEX28 family members were transfected into U2OS cells, TMCC3 specifically localized at the three-way junctions in the peripheral ER. TMCC3 bound to atlastins through the C-terminal transmembrane domains. A TMCC3 mutant lacking the N-terminal coiled-coil domain abolished localization to the three-way junctions, suggesting that TMCC3 localized independently of binding to atlastins. TMCC3 knockdown caused a decrease in the number of three-way junctions and expansion of ER sheets, leading to a reduction of the tubular ER network in U2OS cells. The TMCC3 knockdown phenotype was partially rescued by the overexpression of atlastin-2, suggesting that TMCC3 knockdown would decrease the activity of atlastins. These results indicate that TMCC3 localizes at the three-way junctions for the proper tubular ER network.

3D Crystallization and Structural Studies of Integral Membrane Proteins in Lipidic Cubic phases

2000 ◽  
Vol 56 (s1) ◽  
pp. s83-s83
Author(s):  
P. Nollert ◽  
M. L. Chiu ◽  
M. C. Loewen ◽  
A. Royant ◽  
H. Behrhali ◽  
...  
Keyword(s):  
Membrane Proteins ◽  
Integral Membrane Proteins ◽  
Structural Studies ◽  
Cubic Phases

scholarly journals Automation of large scale transient protein expression in mammalian cells

2011 ◽  
Vol 175 (2) ◽  
pp. 209-215 ◽  
Author(s):  
Yuguang Zhao ◽  
Benjamin Bishop ◽  
Jordan E. Clay ◽  
Weixian Lu ◽  
Margaret Jones ◽  
...  
Keyword(s):  
Protein Expression ◽  
Mammalian Cells ◽  
Large Scale ◽  
Transient Protein Expression ◽  
Expression In Mammalian Cells

A modification of the split-tobacco etch virus method for monitoring interactions between membrane proteins in mammalian cells

2012 ◽  
Vol 423 (1) ◽  
pp. 109-118 ◽  
Author(s):  
Xavier Capdevila-Nortes ◽  
Tania López-Hernández ◽  
Francisco Ciruela ◽  
Raúl Estévez
Keyword(s):  
Membrane Proteins ◽  
Mammalian Cells ◽  
Tobacco Etch Virus

Large scale characterization of plant plasma membrane proteins

Biochimie ◽  
1999 ◽  
Vol 81 (6) ◽  
pp. 655-661 ◽  
Author(s):  
Véronique Santoni ◽  
Patrick Doumas ◽  
David Rouquié ◽  
Monique Mansion ◽  
Thierry Rabilloud ◽  
...  
Keyword(s):  
Plasma Membrane ◽  
Membrane Proteins ◽  
Large Scale ◽  
Plasma Membrane Proteins ◽  
Scale Characterization ◽  
Plant Plasma Membrane

scholarly journals Transgenic goats producing an improved version of cetuximab in milk

2020 ◽  
Author(s):  
Götz Laible ◽  
Sally Cole ◽  
Brigid Brophy ◽  
Paul Maclean ◽  
Li How Chen ◽  
...  
Keyword(s):  
Mammalian Cells ◽  
Large Scale ◽  
Growth Factor Receptor ◽  
Cost Effective ◽  
Scale Production ◽  
Cancer Treatments ◽  
Large Scale Production ◽  
Immunogenic Epitope ◽  
Dependent Cell ◽  
Transgenic Goats

ABSTRACTTherapeutic monoclonal antibodies (mAbs) represent one of the most important classes of pharmaceutical proteins to treat human diseases. Most are produced in cultured mammalian cells which is expensive, limiting their availability. Goats, striking a good balance between a relatively short generation time and copious milk yield, present an alternative platform for the cost-effective, flexible, large-scale production of therapeutic mAbs. Here, we focused on cetuximab, a mAb against epidermal growth factor receptor, that is commercially produced under the brand name Erbitux and approved for anti-cancer treatments. We generated several transgenic goat lines that produce cetuximab in their milk. Two lines were selected for detailed characterization. Both showed stable genotypes and cetuximab production levels of up to 10g/L. The mAb could be readily purified and showed improved characteristics compared to Erbitux. The goat-produced cetuximab (gCetuximab) lacked a highly immunogenic epitope that is part of Erbitux. Moreover, it showed enhanced binding to CD16 and increased antibody-dependent cell-dependent cytotoxicity compared to Erbitux. This indicates that these goats produce an improved cetuximab version with the potential for enhanced effectiveness and better safety profile compared to treatments with Erbitux. In addition, our study validates transgenic goats as an excellent platform for large-scale production of therapeutic mAbs.

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