scholarly journals A Guide to Transient Expression of Membrane Proteins in HEK-293 Cells for Functional Characterization

2016 ◽  
Vol 7 ◽  
Author(s):  
Amanda Ooi ◽  
Aloysius Wong ◽  
Luke Esau ◽  
Fouad Lemtiri-Chlieh ◽  
Chris Gehring
Keyword(s):  
Membrane Proteins ◽  
Transient Expression ◽  
Functional Characterization ◽  
Hek 293 ◽  
Hek 293 Cells ◽  
293 Cells

Functional Characterization of the Regulatory Region of Human CD2-Associated Protein Promoter in HEK 293 Cells

2009 ◽  
Vol 29 (3) ◽  
pp. 203-212 ◽  
Author(s):  
Xin-Ming Su ◽  
Wei Ren ◽  
Chao Lu ◽  
Ji-Qing Chen ◽  
Sheng-Hua Wu ◽  
...  
Keyword(s):  
Functional Characterization ◽  
Regulatory Region ◽  
Hek 293 ◽  
Hek 293 Cells ◽  
293 Cells

Transient expression of osteopontin in HEK 293 cells in serum-free culture

2007 ◽  
Vol 41 (1-2) ◽  
pp. 133-140 ◽  
Author(s):  
Xiangzong Han ◽  
Lei Sun ◽  
Qiangyi Fang ◽  
Dongxiao Li ◽  
Xianghui Gong ◽  
...  
Keyword(s):  
Transient Expression ◽  
Hek 293 ◽  
Serum Free ◽  
Hek 293 Cells ◽  
293 Cells

scholarly journals SLC26 anion exchangers of guinea pig pancreatic duct: molecular cloning and functional characterization

2011 ◽  
Vol 301 (2) ◽  
pp. C289-C303 ◽  
Author(s):  
Andrew K. Stewart ◽  
Boris E. Shmukler ◽  
David H. Vandorpe ◽  
Fabian Reimold ◽  
John F. Heneghan ◽  
...  
Keyword(s):  
Guinea Pig ◽  
Pancreatic Duct ◽  
Xenopus Oocytes ◽  
Functional Characterization ◽  
Pancreatic Ducts ◽  
Anion Exchangers ◽  
Hek 293 ◽  
Hek 293 Cells ◽  
Anion Transporters ◽  
293 Cells

The secretin-stimulated human pancreatic duct secretes HCO3−-rich fluid essential for normal digestion. Optimal stimulation of pancreatic HCO3− secretion likely requires coupled activities of the cystic fibrosis transmembrane regulator (CFTR) anion channel and apical SLC26 Cl−/HCO3− exchangers. However, whereas stimulated human and guinea pig pancreatic ducts secrete ∼140 mM HCO3− or more, mouse and rat ducts secrete ∼40–70 mM HCO3−. Moreover, the axial distribution and physiological roles of SLC26 anion exchangers in pancreatic duct secretory processes remain controversial and may vary among mammalian species. Thus the property of high HCO3− secretion shared by human and guinea pig pancreatic ducts prompted us to clone from guinea pig pancreatic duct cDNAs encoding Slc26a3, Slc26a6, and Slc26a11 polypeptides. We then functionally characterized these anion transporters in Xenopus oocytes and human embryonic kidney (HEK) 293 cells. In Xenopus oocytes, gpSlc26a3 mediated only Cl−/Cl− exchange and electroneutral Cl−/HCO3− exchange. gpSlc26a6 in Xenopus oocytes mediated Cl−/Cl− exchange and bidirectional exchange of Cl− for oxalate and sulfate, but Cl−/HCO3− exchange was detected only in HEK 293 cells. gpSlc26a11 in Xenopus oocytes exhibited pH-dependent Cl−, oxalate, and sulfate transport but no detectable Cl−/HCO3− exchange. The three gpSlc26 anion transporters exhibited distinct pharmacological profiles of 36Cl− influx, including partial sensitivity to CFTR inhibitors Inh-172 and GlyH101, but only Slc26a11 was inhibited by PPQ-102. This first molecular and functional assessment of recombinant SLC26 anion transporters from guinea pig pancreatic duct enhances our understanding of pancreatic HCO3− secretion in species that share a high HCO3− secretory output.

Transient expression of recombinant ACKR4 (CCRL1) gene, an atypical chemokine receptor in human embryonic kidney (HEK 293) cells

2016 ◽  
Vol 43 (7) ◽  
pp. 583-589 ◽  
Author(s):  
Bahareh Parsi ◽  
Abolghasem Esmaeili ◽  
Mohammad Hashemi ◽  
Mohaddeseh Behjati
Keyword(s):  
Transient Expression ◽  
Chemokine Receptor ◽  
Human Embryonic Kidney ◽  
Hek 293 ◽  
Hek 293 Cells ◽  
293 Cells

Functional characterization of rat ρ2 subunits expressed in HEK 293 cells

2005 ◽  
Vol 21 (3) ◽  
pp. 692-700 ◽  
Author(s):  
Anniina Alakuijala ◽  
Katri TalviOja ◽  
Arja Pasternack ◽  
Michael Pasternack
Keyword(s):  
Functional Characterization ◽  
Hek 293 ◽  
Hek 293 Cells ◽  
293 Cells

scholarly journals Transient Expression in HEK 293 Cells: An Alternative to E. coli for the Production of Secreted and Intracellular Mammalian Proteins

2014 ◽  
pp. 209-222 ◽  
Author(s):  
Joanne E. Nettleship ◽  
Peter J. Watson ◽  
Nahid Rahman-Huq ◽  
Louise Fairall ◽  
Mareike G. Posner ◽  
...  
Keyword(s):  
Transient Expression ◽  
Hek 293 ◽  
E Coli ◽  
Hek 293 Cells ◽  
Mammalian Proteins ◽  
293 Cells

scholarly journals Characteristics and requirements of basal autophagy in HEK 293 cells

Autophagy ◽  
2013 ◽  
Vol 9 (9) ◽  
pp. 1407-1417 ◽  
Author(s):  
Patience Musiwaro ◽  
Matthew Smith ◽  
Maria Manifava ◽  
Simon A. Walker ◽  
Nicholas T. Ktistakis
Keyword(s):  
Hek 293 ◽  
Hek 293 Cells ◽  
293 Cells

scholarly journals Using kICS to Reveal Changed Membrane Diffusion of AQP-9 Treated with Drugs

Membranes ◽  
2021 ◽  
Vol 11 (8) ◽  
pp. 568
Author(s):  
Jakob L. Kure ◽  
Thommie Karlsson ◽  
Camilla B. Andersen ◽  
B. Christoffer Lagerholm ◽  
Vesa Loitto ◽  
...  
Keyword(s):  
Diffusion Coefficient ◽  
Plasma Membrane ◽  
Membrane Proteins ◽  
Actin Polymerization ◽  
Correlation Spectroscopy ◽  
Image Correlation ◽  
Membrane Diffusion ◽  
Hek 293 ◽  
293 Cells ◽  
Aquaporin 9

The formation of nanodomains in the plasma membrane are thought to be part of membrane proteins regulation and signaling. Plasma membrane proteins are often investigated by analyzing the lateral mobility. k-space ICS (kICS) is a powerful image correlation spectroscopy (ICS) technique and a valuable supplement to fluorescence correlation spectroscopy (FCS). Here, we study the diffusion of aquaporin-9 (AQP9) in the plasma membrane, and the effect of different membrane and cytoskeleton affecting drugs, and therefore nanodomain perturbing, using kICS. We measured the diffusion coefficient of AQP9 after addition of these drugs using live cell Total Internal Reflection Fluorescence imaging on HEK-293 cells. The actin polymerization inhibitors Cytochalasin D and Latrunculin A do not affect the diffusion coefficient of AQP9. Methyl-β-Cyclodextrin decreases GFP-AQP9 diffusion coefficient in the plasma membrane. Human epidermal growth factor led to an increase in the diffusion coefficient of AQP9. These findings led to the conclusion that kICS can be used to measure diffusion AQP9, and suggests that the AQP9 is not part of nanodomains.

Halothane Inhibition of Recombinant Cardiac L-type Ca2+ Channels Expressed in HEK-293 Cells

2005 ◽  
Vol 103 (6) ◽  
pp. 1156-1166 ◽  
Author(s):  
Kevin J. Gingrich ◽  
Son Tran ◽  
Igor M. Nikonorov ◽  
Thomas J. Blanck
Keyword(s):  
Charge Transfer ◽  
Calcium Channels ◽  
Dependent Manner ◽  
Current Time ◽  
Hek 293 ◽  
Hek 293 Cells ◽  
293 Cells ◽  
Dependent Inactivation ◽  
Voltage Dependent

Background Volatile anesthetics depress cardiac contractility, which involves inhibition of cardiac L-type calcium channels. To explore the role of voltage-dependent inactivation, the authors analyzed halothane effects on recombinant cardiac L-type calcium channels (alpha1Cbeta2a and alpha1Cbeta2aalpha2/delta1), which differ by the alpha2/delta1 subunit and consequently voltage-dependent inactivation. Methods HEK-293 cells were transiently cotransfected with complementary DNAs encoding alpha1C tagged with green fluorescent protein and beta2a, with and without alpha2/delta1. Halothane effects on macroscopic barium currents were recorded using patch clamp methodology from cells expressing alpha1Cbeta2a and alpha1Cbeta2aalpha2/delta1 as identified by fluorescence microscopy. Results Halothane inhibited peak current (I(peak)) and enhanced apparent inactivation (reported by end pulse current amplitude of 300-ms depolarizations [I300]) in a concentration-dependent manner in both channel types. alpha2/delta1 coexpression shifted relations leftward as reported by the 50% inhibitory concentration of I(peak) and I300/I(peak)for alpha1Cbeta2a (1.8 and 14.5 mm, respectively) and alpha1Cbeta2aalpha2/delta1 (0.74 and 1.36 mm, respectively). Halothane reduced transmembrane charge transfer primarily through I(peak) depression and not by enhancement of macroscopic inactivation for both channels. Conclusions The results indicate that phenotypic features arising from alpha2/delta1 coexpression play a key role in halothane inhibition of cardiac L-type calcium channels. These features included marked effects on I(peak) inhibition, which is the principal determinant of charge transfer reductions. I(peak) depression arises primarily from transitions to nonactivatable states at resting membrane potentials. The findings point to the importance of halothane interactions with states present at resting membrane potential and discount the role of inactivation apparent in current time courses in determining transmembrane charge transfer.

Micropatterned surfaces of PDMS as growth templates for HEK 293 cells

2007 ◽  
Vol 9 (4) ◽  
pp. 475-485 ◽  
Author(s):  
R. M. Johann ◽  
Ch. Baiotto ◽  
Ph. Renaud
Keyword(s):  
Hek 293 ◽  
Hek 293 Cells ◽  
293 Cells
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