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

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

Characterization of glycine release mediated by glycine transporter 1 stably expressed in HEK-293 cells

1997 ◽  
Vol 49 (1-2) ◽  
pp. 89-94 ◽  
Author(s):  
Kazuko Sakata ◽  
Kohji Sato ◽  
Patrick Schloss ◽  
Heinrich Betz ◽  
Shoichi Shimada ◽  
...  
Keyword(s):  
Hek 293 ◽  
Hek 293 Cells ◽  
Glycine Release ◽  
Glycine Transporter 1 ◽  
Glycine Transporter ◽  
293 Cells

Cloning, expression, and functional characterization of the von Willebrand factor–cleaving protease (ADAMTS13)

Blood ◽  
2002 ◽  
Vol 100 (10) ◽  
pp. 3626-3632 ◽  
Author(s):  
Barbara Plaimauer ◽  
Klaus Zimmermann ◽  
Dirk Völkel ◽  
Gerhard Antoine ◽  
Randolf Kerschbaumer ◽  
...  
Keyword(s):  
Von Willebrand Factor ◽  
Functional Activity ◽  
Functional Characterization ◽  
Eukaryotic Expression ◽  
Gene Encoding ◽  
Hek 293 ◽  
293 Cells ◽  
Von Willebrand ◽  
Willebrand Factor

Deficient von Willebrand factor (VWF) degradation has been associated with thrombotic thrombocytopenic purpura (TTP). In hereditary TTP, the specific VWF-cleaving protease (VWF-cp) is absent or functionally defective, whereas in the nonfamilial, acquired form of TTP, an autoantibody inhibiting VWF-cp activity is found transiently in most patients. The gene encoding for VWF-cp has recently been identified as a member of the metalloprotease family and designatedADAMTS13, but the functional activity of the ADAMTS13 gene product has not been verified. To establish the functional activity of recombinant VWF-cp, we cloned the complete cDNA sequence in a eukaryotic expression vector and transiently expressed the encoded recombinant ADAMTS13 in HEK 293 cells. The expressed protein degraded VWF multimers and proteolytically cleaved VWF to the same fragments as those generated by plasma VWF-cp. Furthermore, recombinant ADAMTS13-mediated degradation of VWF multimers was entirely inhibited in the presence of plasma from a patient with acquired TTP. These data show that ADAMTS13 is responsible for the physiologic proteolytic degradation of VWF multimers.

scholarly journals Characterization of a human 5-hydroxytryptamine3 receptor type A (h5-HT3R-AS) subunit stably expressed in HEK 293 cells

1996 ◽  
Vol 118 (5) ◽  
pp. 1237-1245 ◽  
Author(s):  
Anthony G. Hope ◽  
John A. Peters ◽  
Angus M. Brown ◽  
Jeremy J. Lambert ◽  
Thomas P. Blackburn
Keyword(s):  
Type A ◽  
Receptor Type ◽  
Hek 293 ◽  
Hek 293 Cells ◽  
293 Cells

scholarly journals Characterization of bovine interleukin-2 stably expressed in HEK-293 cells

2020 ◽  
Author(s):  
Shuya MITOMA ◽  
Heba M. EL-KHAIAT ◽  
Tomofumi UTO ◽  
Katsuaki SATO ◽  
Satoshi SEKIGUCHI ◽  
...  
Keyword(s):  
Interleukin 2 ◽  
Hek 293 ◽  
Hek 293 Cells ◽  
293 Cells

scholarly journals Clinical and Functional Characterization of the Recurrent TUBA1A p.(Arg2His) Mutation

2018 ◽  
Vol 8 (8) ◽  
pp. 145 ◽  
Author(s):  
Jennifer Gardner ◽  
Thomas Cushion ◽  
Georgios Niotakis ◽  
Heather Olson ◽  
P. Grant ◽  
...  
Keyword(s):  
De Novo ◽  
Functional Characterization ◽  
Clinical Manifestations ◽  
Fetal Brain ◽  
Detailed Comparison ◽  
Hek 293 ◽  
293 Cells ◽  
Brain Malformations ◽  
Computer Based

The TUBA1A gene encodes tubulin alpha-1A, a protein that is highly expressed in the fetal brain. Alpha- and beta-tubulin subunits form dimers, which then co-assemble into microtubule polymers: dynamic, scaffold-like structures that perform key functions during neurogenesis, neuronal migration, and cortical organisation. Mutations in TUBA1A have been reported to cause a range of brain malformations. We describe four unrelated patients with the same de novo missense mutation in TUBA1A, c.5G>A, p.(Arg2His), as found by next generation sequencing. Detailed comparison revealed similar brain phenotypes with mild variability. Shared features included developmental delay, microcephaly, hypoplasia of the cerebellar vermis, dysplasia or thinning of the corpus callosum, small pons, and dysmorphic basal ganglia. Two of the patients had bilateral perisylvian polymicrogyria. We examined the effects of the p.(Arg2His) mutation by computer-based protein structure modelling and heterologous expression in HEK-293 cells. The results suggest the mutation subtly impairs microtubule function, potentially by affecting inter-dimer interaction. Based on its sequence context, c.5G>A is likely to be a common recurrent mutation. We propose that the subtle functional effects of p.(Arg2His) may allow for other factors (such as genetic background or environmental conditions) to influence phenotypic outcome, thus explaining the mild variability in clinical manifestations.

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.

scholarly journals Molecular cloning and functional characterization of KCC3, a new K-Cl cotransporter

1999 ◽  
Vol 277 (6) ◽  
pp. C1210-C1219 ◽  
Author(s):  
Joanne E. Race ◽  
Fadi N. Makhlouf ◽  
Paul J. Logue ◽  
Frederick H. Wilson ◽  
Philip B. Dunham ◽  
...  
Keyword(s):  
Functional Characterization ◽  
Amino Acid Level ◽  
Transmembrane Domains ◽  
Chromosome 15 ◽  
Hek 293 ◽  
293 Cells ◽  
Liver And Pancreas ◽  
Large Extracellular Loop ◽  
Hydropathy Analysis

We isolated and characterized a novel K-Cl cotransporter, KCC3, from human placenta. The deduced protein contains 1,150 amino acids. KCC3 shares 75–76% identity at the amino acid level with human, pig, rat, and rabbit KCC1 and 67% identity with rat KCC2. KCC3 is 40 and 33% identical to two Caenorhabditis elegans K-Cl cotransporters and ∼20% identical to other members of the cation-chloride cotransporter family (CCC), two Na-K-Cl cotransporters (NKCC1, NKCC2), and the Na-Cl cotransporter (NCC). Hydropathy analysis indicates a typical KCC topology with 12 transmembrane domains, a large extracellular loop between transmembrane domains 5 and 6 (unique to KCCs), and large NH2 and COOH termini. KCC3 is predominantly expressed in kidney, heart, and brain, and is also expressed in skeletal muscle, placenta, lung, liver, and pancreas. KCC3 was localized to chromosome 15. KCC3 transiently expressed in human embryonic kidney (HEK)-293 cells fulfilled three criteria for increased expression of K-Cl cotransport: stimulation of cotransport by swelling, treatment with N-ethylmaleimide, or treatment with staurosporine.

scholarly journals Genomic identification and biochemical characterization of the mammalian polyamine oxidase involved in polyamine back-conversion

2003 ◽  
Vol 370 (1) ◽  
pp. 19-28 ◽  
Author(s):  
Slavoljub VUJCIC ◽  
Ping LIANG ◽  
Paula DIEGELMAN ◽  
Debora L. KRAMER ◽  
Carl W. PORTER
Keyword(s):  
Northern Blot Analysis ◽  
Biochemical Characterization ◽  
Polyamine Oxidase ◽  
Preference Ranking ◽  
Hek 293 ◽  
Hek 293 Cells ◽  
Polyamine Analogues ◽  
293 Cells ◽  
Back Conversion

In the polyamine back-conversion pathway, spermine and spermidine are first acetylated by spermidine/spermine N1-acetyltransferase (SSAT) and then oxidized by polyamine oxidase (PAO) to produce spermidine and putrescine respectively. Although PAO was first purified more than two decades ago, the protein has not yet been linked to genomic sequences. In the present study, we apply a BLAST search strategy to identify novel oxidase sequences located on human chromosome 10 and mouse chromosome 7. Homologous mammalian cDNAs derived from human brain and mouse mammary tumour were deduced to encode proteins of approx. 55kDa having 82% sequence identity. When either cDNA was transiently transfected into HEK-293 cells, intracellular spermine pools decreased by approx. 30%, whereas spermidine increased 2—4-fold. Lysates of human PAO cDNA-transfected HEK-293 cells, but not vector-transfected cells, rapidly oxidized N1-acetylspermine to spermidine. Substrate specificity determinations with the lysate assay revealed a preference ranking of N1-acetylspermine = N1-acetylspermidine>N1,N12-diacetylspermine>spermine; spermidine was not acted upon. This ranking is identical to that reported for purified PAO and distinctly different from the recently identified spermine oxidase (SMO), which prefers spermine over N1-acetylspermine. Monoethyl- and diethylspermine analogues also served as substrates for PAO, and were internally cleaved adjacent to a secondary amine. We deduce that the present oxidase sequences are those of the FAD-dependent PAO involved in the polyamine back-conversion pathway. In Northern blot analysis, PAO mRNA was much less abundant in HEK-293 cells than SMO or SSAT mRNA, and all three were differentially induced in a similar manner by selected polyamine analogues. The identification of PAO sequences, together with the recently identified SMO sequences, provides new opportunities for understanding the dynamics of polyamine homoeostasis and for interpreting metabolic and cellular responses to clinically-relevant polyamine analogues and inhibitors.

Pharmacological characterization of new β-agonists using Huβ1- and Huβ2-adrenergic receptor binding assay in transfected HEK-293 cells

2005 ◽  
Vol 529 (1-2) ◽  
pp. 53-56 ◽  
Author(s):  
Cinzia Civitareale ◽  
Caterina Ambrosio ◽  
Maria Sbraccia ◽  
Maurizio Fiori ◽  
Gianfranco Brambilla ◽  
...  
Keyword(s):  
Receptor Binding ◽  
Adrenergic Receptor ◽  
Binding Assay ◽  
Pharmacological Characterization ◽  
Hek 293 ◽  
Hek 293 Cells ◽  
Receptor Binding Assay ◽  
293 Cells
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